DEHYDRATED CULTURE MEDIA 



121 



Preparation of Media 



Prepare a double strength Proteose No. 3 Agar by suspending 9.0 grams in 

 100 ml. of cold distilled water, and steaming or boiling to dissolve the medium 

 completely. At the same time prepare a double strength hemoglobin solution by 

 placing 2 grams Bacto-Hemoglobin in a dry flask and adding 100 ml. cold dis- 

 tilled water, while the flask is being agitated vigorously. The Bacto-Hemoglobin 

 suspension is shaken intermittently for 10-15 minutes to break up all aggregates 

 and effect complete solution. Both double strength solutions of Proteose No. 3 

 Agar and Bacto-Hemoglobin are sterilized in the autoclave for 15 minutes at 

 15 pounds pressure (121°C.). 



The sterile solutions are allowed to cool to 50-60° C. Add 1 per cent of Bacto- 

 Supplement A or Bacto-Supplement B, based on the final volume of medium, to 

 the double strength Proteose No. 3 Agar (2 ml. per 100 ml. double strength 

 sterile agar base). Mix well and add an equal volume of double strength sterile 

 Bacto-Hemoglobin solution at 50-60° G. Mix well and distribute into sterile petri 

 dishes. Caution: Do not add Bacto-Supplement A or Bacto-Supplement B to the 

 hemoglobin solution before mixing with the agar base. Best results will be ob- 

 tained if the medium is prepared in advance so that the surface of the medium 

 is dry before inoculation. 



When the single strength medium, without enrichment, is desired, the medium 

 is rehydrated by suspending 45 grams of Bacto-Proteose No. 3 Agar in 1000 ml. 

 of cold distilled water and heating to boiling to dissolve the medium completely. 

 Distribute in tubes or flasks and sterilize in the autoclave for 15 minutes at 15 

 pounds pressure (121°C.). The final reaction of the medium without enrich- 

 ment will be pH 7.3. 



One pound of Bacto-Proteose No. 3 Agar will make 10 liters of medium. 



Acknowledgment 



We are indebted to the personnel of the Division of Social Hygiene of the 

 Detroit Department of Health for their cooperation in supplying the clinical 

 material for these studies. 



1 Am. J. Syphilis Gonorrh. Venereal Diseases, 

 33:164:1949. 



2 Muench. Wochschr., 80:846:1933. 

 8 J. Path. Bact., 39:221:1934. 



* J. Infectious Diseases, 61:129:1937. 

 ^ Am. J. Syphilis, 20:347:1936. 

 «Am. J. Syphilis, 22:55:1938. 

 'Seventh Annual Yearbook (1936-37), 



p. 133, Suppl., Am. J. Pub. Health, 

 27:No. 3:1937. 

 ^ Bull. Genitoinfectious Diseases, Mass. State 

 Health Dept., 2:1:1938. 



* Pelouze : Gonorrhea in the Male and 

 Female, 3rd Edition: 55: 1939. 



^0 Am. J. Syphilis Gonorrh. Venereal Diseases, 



and 

 3rd 



25:22:1941. 

 ^ Am. J. Public Health, 31:457:1941, 



Venereal Disease Inform., 

 Venereal Disease Inform., 

 Bact., 44:139:1942. 

 Bact., 45:321:1943. 

 Bact., 45:410:1943. 



24:218:1943. 

 24:133:1943. 



" Personal Communication 1943. 

 ^ Personal Communication 1944. 

 "^ Am. J. Syphilis Gonorrh. Venereal Diseases, 

 28:634:1944. 



20 U.S. Naval Med. Bull., 43:409:1944. 



21 Encyclopedia of Medicine, Surgery 

 Specialties, 3rd Edition. Now in press. 



22 Diagnostic Procedures and Reagents, 

 Edition: 107: 1950. 



23 J. Bact., 40:162:1940. 

 2* Am. J. Syphilis Gonorrh. Venereal Diseases, 



29:19:1945- 



25 J. Bact., 50:589:1945. 



26 Am. J. Clin. Path., 18:256:1948. 

 2^ J. Venereal Disease Inform., 23:224:1942. 

 2S J. Lab. Clin. Med., 27:656:1942. 



29 Proc. Soc. Exp. Biol. Med., 48:330:1941. 



30 J. Am. Med. Assoc, 123:547:1943. 



31 Personal Communication 1945. 



^ Am. J. Syphilis Gonorrh. Venereal Diseases, 

 28:397:1944. 



