784 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



potassium sulphocyanide. (CaCl2 

 may be added also if desired). 



(4) Adjust to faint alkalinity. 



(5) Distribute into test tubes. 



(6) Sterilize and coagulate (method not 

 specified). 



Coplans reported that B. diphtheriae 

 colonies after 18 hours were pink 

 and a bluish pink tint diffused out 

 into the medium. Medium originally 

 yellow. 1.0% KCNS sufficed for this 

 purpose. If 2.0% be added B. Hof- 

 manni inhibited. If more than 2\% 

 be added B. diphtheria and Torulae 

 are inhibited. 



(b) Coplans used 0.5 to 2.0% ofPotas- 

 sium-ferri-cyanide instead of 1.25 g. 

 of potassium-sulpho-cyanide as in 

 variant above. (0.5 to 2.0% KCNS 

 may be added). 



(c) Coplans used l.Og. potassium-sulpho- 

 cyanide instead of 1.25 g. as in variant 

 (a) above and added 0.5 to 2.0 g. 

 boric acid. (The KCNS may be 

 omitted.) 



(d) Coplans added 1.0 g. Potassium 

 ferrocyanide instead of 1.0% potas- 

 sium sulpho-cyanide as in variant 

 (a). (1.0 g. potassium sulpho-cy- 

 anide may also be added if desired). 



(e) Harvey added 0.5% glucose, 1.0% 

 potassium sulpho-cyanide, and 0.5% 

 of a 2.0% neutral red solution to 

 infusion broth, see variant (bb) 779. 

 One part of this mixture was added 

 to 3 parts sheep serum and co- 

 agulated. 



References: Rankin (1911 p. 273), Coplans 

 (1911 pp. 287, 282), Harvey (1921-22 

 p. 81). 



2419. Ball's Glucose Bouillon Blood Albumin 

 Medium 



Constituents : 



1. Glucose bouillon 85.0 cc. 



2. Blood albumin (commercial).. 15.0 g. 

 Preparation : 



(1) Prepare glucose bouillon. 



(2) Dissolve 15 parts commercial blood 

 albumin in 85 parts (1). 



(3) Tube. 



(4) Inspissate in a slanting position. 

 Sterilization: Remove the tubes and ster- 



ilize by the fractional method in the 



steamer. 

 Use: General culture medium. 

 Reference: Ball (1919 p. 84). 



2420. Crowe's Glucose Agar Blood Medium 

 Constituents : 



1. Nutrient agar 250.0 cc. 



2. Blood, defibrinated (bullock). 750.0 cc. 



3. Glucose (1.0%) iq.q g. 



Preparation : 



(1) Strain about a liter of defibrinated 

 bullock blood thru muslin. 



(2) Place 750.0 cc. of (1) in a water bath 

 at 50°C. 



(3) Melt 250.0 cc. of peptone (or trypsin) 

 agar of the usual reaction. 



(4) Mix (2) and (3) and add 1.0% glucose. 



(5) Keep at 50°C. while plates or tubes 

 are filled. At least 0.25 of an inch is 

 required for each plate. 



(6) Place in a steam sterilizer at 65 to 

 70°C. for an hour or two and then at 

 80-90°C. on each of 3 successive days. 



Sterilization: See step (6) above. 

 Use: Differentiation of streptococci. 

 Reference: Crowe (1921 p. 486). 



2421. Vardon's Bouillon Serum 

 (Desiccated) 

 Constituents : 



1. Bouillon 100.0 cc. 



2. Horse serum 300.0 cc. 



Preparation : 



(1) Mix 1 part bouillon with 3 parts 

 serum. 



(2) Pour the mi.xture into shallow trays. 



(3) Dry at a temperature not e.xceeding 

 50°C. 



(4) Scrape the dried film off the trays and 

 store for use. 



(5) Dissolve the required amount of (4) 

 in cold water. (Amount required to 

 be calculated from the amount of 

 serum and bouillon started with and 

 amount of powder obtained). 



(6) Distribute when dissolved in amounts 

 of about 7.0 cc. into test tubes. 



(7) Inspissate the medium in a sloping 

 position at a temperature of 75°C. 

 for 60 minutes on three successive 

 days. 



(8) Incubate tubes 24 hours and reject 

 any which are not sterile. 



