508 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



(9) Filter thru filter paper. 



(10) Dissolve 3.0% agar in (9) in the 

 steamer. 



(11) Adjust the reaction by the addition 

 of NaOH so that 1.3% NaOH will 

 be required to bring the reaction 

 to the neutral point of phenol- 

 phthalein. 



(12) Heat. 



(13) Filter in the steamer. 



(14) Distribute in 100.0 cc. lots in Erlen- 

 meyer flasks. 



(15) Prepare a 0.2% china green solution. 



(16) Add 1.4 to 1.5 cc. of (15) to each 

 100.0 cc. of sterile melted and cooled 

 to 60-65°C. (14). Mix well. 



(17) Pour into plates. 

 Sterilization: Method of sterilization of 



(14) not given. 



Use: Isolation of typhoid bacilli from 

 feces. Enrichment of typhoid bacilli. 

 Smear the plates with the stool or feces 

 suspension. Incubate 20 hours at 37°C. 

 Wash the surface of the plates with 8 to 

 10.0 cc. physiological salt solution, and 

 smear 1 to 3 loops of the salt solution sus- 

 pension on Drigalski-Conradi medium. 



Reference: Werbitzki (1909 p. 205). 



1682. league's Victoria Blue Infusion Agar 



Constituents : 



1. Meat infusion 1000.0 cc. 



2. Peptone, Witte (1.0%) 10.0 g. 



3. NaCl (0.5%) 5.0 g. 



4. Victoria blue 4R 



Preparation : 



(1) Detailed method of preparation or 

 composition of meat infusion agar 

 not given except that it contains 1.0% 

 Witte's peptone and 0.5% NaCl, and 

 that it is to be prepared in the usual 

 manner using the Arnold sterilizer. 



(2) Clear (1) with egg white. 



(3) Filter thru cotton. 



(4) Titrate to +1- 



(5) Add stock solutions (method of 

 preparation not given) of Victoria 

 blue 4R in varying amounts so that 

 there is present 1/20, 1/30, 1/40 or 1/50 

 of dye, to sterile (4). 



Sterilization: Sterilize by heating in Ar- 

 nold for 3 successive days. 



Use: Enrichment of B. paratijphosus A and 

 B. enteritidis. Author reported that the 



dye inhibited growth of B. paratyphoid B 

 but did not hinder growth of B. paraty- 

 phoid A or B. enteritidis . 

 Reference: Teague (1918 p. 1). 



1683. Drennan and Teague's Crystal Violet 

 Infusion Agar 



Constituents : 



1. Distilled water 1000.0 cc. 



2. Heart, beef 1.0 lb. 



3. Peptone 10.0 g. 



4. NaCl 2.5 g. 



5. Agar 15.0 g. 



6. N/1 NaOH 4.0 cc. 



7. Crystal violet 1/70 g. 



Preparation: 



(1) Pass one pound of beef heart thru a 

 meat chopping machine and soak in 

 one liter of distilled water over night 

 in an ice box. 



(2) Squeeze the fluid thru a cheese cloth, 

 heat to boiling and filter thru filter 

 paper. 



(3) Add 1.0% peptone, 0.25% NaCl, 

 1.5% agar-agar, 4.0 cc. of N/1 NaOH 

 and heat for 30 minutes in the auto- 

 clave at 15 pounds pressure. 



(4) Adjust the reaction to -fl (hot titra- 

 tion), clear with white of egg. 



(5) Filter thru cotton. 



(6) Distribute in flasks in 200.0 cc. lots. 



(7) Add to the melted sterile agar 1/700% 

 of crystal violet. 



(8) Pour in plates. 



Sterilization: Sterilize in the autoclave for 

 20 minutes at 15 pounds pressure. 



Use: Isolation of B. pestis from lesions. 



Reference: Drennan and Teague (1919 

 pp. 521-529). 



1684. Meyer and Batchelder's Sulphite 

 Gentian Violet Infusion Agar 



Constituents : 



1. Infusion agar 1000.0 cc. 



2. NaoSOa (10.0% soln.) 2.5 cc. 



3. Gentian violet (1 : 1000) 25.0 cc. 



Preparation : 



(1) Prepare heart infusion agar using 

 Berna peptone, according to Hun- 

 toon's method. (See medium 1863 

 for Huntoon's method.) 



(2) Add 0.25% of a freshly prepared 10.0% 

 solution) NasSOa and 1/400% (2.5 cc. 

 of a 1:1000 solution per 100.0 cc.) of 



