CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



531 



(1), keeping the temperature below 

 60°C. 



(8) Titrate and adjust the reaction to 

 neutral to phenolphthalein adding 

 normal HCl or NaOH. 



(9) Heat on a water bath for 40 min- 

 utes. 



(10) Make up lost weight. 



(11) Readjust to neutrality if neces- 

 sary and boil 5 minutes. 



(12) Restore lost weight. 



(13) Filter thru absorbent cotton and 

 cotton flannel. 



(14) Titrate and record final reaction. 



(15) Add 1.0% lactose and sufficient 

 azolitmin solution. 



(16) Tube in 10.0 cc. quantities. 



(17) Sterilize for 15 minutes in the 

 autoclave at 120°C. or for 30 min- 

 utes on each 3 successive days. 



(d) Giltner gave the following methods 



of preparation. The medium was 



used in water analysis. 



I. (1) Prepare infusion agar using 



equal parts meat infusion and 



water, with 1.0% peptone and 



1.5% agar. 



(2) Adjust the reaction to -f-1.0. 



(3) Add 1.0% lactose and 2.0% 

 azolitmin just before tubing. 



(4) Tube. 



(5) Sterilize for 30 minutes on 3 

 successive days. 



II. (1) Preparation of meat infusion 

 not given. 



(2) Strain (1) thru a piece of clean 

 cheese cloth. 



(3) Place 2.0% washed agar in 

 distilled water. 



(4) Weigh (3). 



(5) Digest over a free flame. 



(6) Add distilled water to make 

 up the loss in weight. 



(7) Add lactose 2.0% and peptone 

 2.0% to (6) and mix until solu- 

 tion is complete. 



(8) Add (2) to (7). 



(9) Adjust the reaction to 0. 



(10) Add 2.0% azolitmin solution. 



(11) Boil over a free flame. 



(12) Distribute in 100.0 cc. lots in 

 250.0 cc. Florence flasks. 



(13) Sterilization not specified. 



References: Smith (1902 p. 94), (1905, 

 p. 196), Heineman (1905 p. 127), Com- 

 mittee A. P. H. A. (1913 p. 129), Giltner 

 (1921 p. 379, 380). 



1731. Bitter's China Blue Malachite Green 

 Agar 



Constituents : 



1. Infusion agar (containing 

 2.0 or 3.0% peptone and 



NaCl) 1000.0 cc. 



2. Lactose 20.0 g. 



3. China blue (sat. aqueous 

 solution (Hochster)) 90 drops 



4. Malachite green (0.1% 

 solution, crystalline extra 

 Hochst.) 25.0 cc. 



Preparation : 



(1) Prepare meat infusion agar with 

 2.0 or 3.0% peptone and NaCl. 



(2) Neutralize (1) with N/1 NaOH. 



(3) Add 2.0% lactose. 



(4) Boil a few minutes. 



(5) To each 100.0 cc. of (4) add 9 drops 

 of a saturated (about 10%) watery 

 china blue solution (Hochster Farb- 

 werks 10 g. 2M) by means of a 1.0 cc. 

 pipette. 



(6) Then add to each 100.0 cc. of (5) 

 2,5 cc. of a 0.1% malachite green 

 (cryst extra Hochst, ) solution. 



(7) Pour sterile (6) in thin layers in 

 sterile plates. 



Sterilization: Sterilize 10 minutes in the 

 autoclave. 



Use: Diagnosis of typhoid fever. Author 

 reported that all acid forming organisms 

 (coli, etc.) gave bright blue colonies. 

 All non-lactose fermenting organisms 

 (typhoid) or alkali forming organisms 

 gave a colorless or yellowish colony. 



Reference: Bitter (1911 p. 474). 



1732. Guth's Alizarine Lactose Agar 

 (Klimmer) 



Constituents : 



1. Meat infusion (beef or 



horse) 1000.0 cc. 



2. Agar 30.0 g. 



3. Peptone 10.0 g. 



4. NaCl 5.0 g. 



5. Lactose 10.0 g. 



6. Alizarin 0.8 g. 



