620 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



water solution of congo red to each 

 flask. 

 Sterilization: Method not given. 

 Use; Water analysis. Detection of B. coli. 

 Author reported that B. coli colonies were 

 intensive black (bluish black). B. typhi, 

 B. paratyphi A and B, B. dysentery, 

 Shiga Flexner Y, B. enteritidis, B. 

 cholerae did not show this black or dark 

 color. B. acidi lactici and B. lactis- 

 aerogenes gave the same reaction as 

 B. coli. 

 Reference: Galli-Valerio (1916 p. 136). 



1938. Teague and Travis' Eosin Bismark 

 Brown Nutrose Agar 



Constituents: 



1. Distilled water 2000.0 cc. 



2. Chopped beef 2.0 lbs. 



3. Peptone (Witte) 20 e 



4- NaCl lO.oJ; 



5. Nutrose 5 g. 



6. Sucrose 20.0 g. 



7. Eosin 3.0% bluish solution . 40.0 cc, 



8. Bismark brown 1.0% solu- 

 tion 80.0 cc. 



9- Agar 15 g. 



Preparation : 



(1) Soak beef in water in ice chest over 

 night. 



(2) Squeeze out fluid. 



(3) Heat liquid in Arnold sterilizer and 

 filter thru filter paper. 



(4) Neutralize with NaOH and heat 

 again . 



(5) Cool and inoculate with B. coli and 

 incubate for 2 or 3 days. 



(6) Add 3, 4 and 9 to (5). 



(7) Clear with egg. Filter. 



(8) Add nutrose. 



(9) Prepare a 3.0% stock solution of 

 bluish eosin in distilled water. 

 Store in the dark. 



(10) Prepare a 1.0% solution of bismark 

 brown in water containing 10.0% 

 alcohol (not soluble to 1.0% in 

 pure water). 



(11) To 50.0 cc. of (8) add 1.0% sucrose 

 (0.5 g.), 1.0 cc. of (9) and 2.0 cc 

 of (10). 



(12) Shake well. 



(13) Pour into sterile Petri dishes. 



(14) Dry by inverting plates in the incu- 

 bator for 20 to 30 minutes. 

 Sterilization: Not specified. 

 Use: To isolate cholera vibrio. Authors 

 reported that cholera vibrio colonies had 

 a dark colored center. Liebig's meat 

 extract may be substituted for infusion. 

 Reference : Teague and Travis (1916 p. 602) . 



1939. Gassner's Nutrose Agar 

 Constituents : 



1. Meat infusion 1000.0 cc. 



2. Peptone jq.O g. ' 



3. Nutrose 10.0 g. 



4- Agar 30 og' 



SNaCl 50g. 



Preparation : 



(1) Prepare meat infusion. 



(2) Dissolve 2, 3, 4 and 5 in 1. 



(3) Adjust to slightly alkaline to litmus. 

 Sterilization: Not specified. 



Use: Cultivation of colon-typhoid group. 

 Reference: Gassner (1916-17 p. 315). 



1940. Loeffler's Malachite Green Nutrose 



Agar (Roddy) 

 Constituents: 



1. Distilled water 500.0 cc. 



2. Infusion broth 500.0 cc. 



3- Agar 30.0 g. 



4. Nutrose lo.O g. 



5. Malachite green 



(2.0% soln.) 20.0 to 30.0 cc. 



Preparation : 



(1) Prepare infusion broth. 



(2) Mix equal volumes of (1) and dis- 

 tilled water. 



(3) Neutralize (indicator not specified). 



(4) Add 7.5 cc. of a normal HCl solution." 



(5) Dissolve 30.0 g. of shredded agar in 

 (4) by boiling. 



(6) Neutralize by the addition of so- 

 dium hydrate. 



(7) Add 5.0 cc. of normal NajCOs solu- 

 tion. 



(8) Steam for 2 hours. 



(9) Add 100.0 cc. of a 10.0% nutrose 

 solution. 



(10) When ready for use, liquefy sterile 

 (9) and add between 2.0 and 3.0 cc. 

 of a 2.0% aqueous solution of mala- 

 chite green (Hochst 120) to each 

 10.0 cc. of medium. 



