CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



575 



Use: Enrichment medium for cholera 



vibrio. 

 Reference: Galli-Valerio (1912 p. 550). 



1842. Padlewsky's Malachite Green Bile 

 Agar 



Constituents : 



1. Meat extract agar 



(3.0%) 1000.0 cc. 



2. Peptone 20.0 g. 



3. Bile 35.0 cc. 



4. Lactose (c.p.) 10.0 g. 



5. Malachite green 

 (1.0% aq. solution) 

 (Hochst) 5.0 cc. 



6. Sodium sulphite 

 (10.0% aq. pur. solu- 

 tion) 7.5 to 10.0 cc. 



Preparation : 



(1) Prepare a 3.0% meat extract agar. 

 (Meat infusion may be used instead 

 of meat extract). 



(2) Treat the ox bile with steam in a 

 Koch's apparatus. (Time not 

 specified). 



(3) Filter the bile thru cotton. 



(4) Add 2.0% peptone, 3.0%, (3) and 

 1.0% c.p. lactose to (1). 



(5) Adjust the reaction to slightly 

 alkaline to litmus. 



(6) It is not necessary to filter the agar 

 after the addition of the filtered bile. 



(7) Distribute into flasks in 100 to 200 cc. 

 lots. 



(8) When ready for use melt the sterile 

 agar, cool to 60 to 65 °C. 



(9) Add to each 100.0 cc. of (8) 0.5 cc. 

 of a 1.0% watery solution of Hochst 

 chemically pure crystalline mala- 

 chite green, 0.5 cc. bile and 0.75 to 

 1.0 cc. of a 10.0% watery solution of 

 sodium sulphite (pur.) by means of a 

 pipette. 



(10) Mix thoroly. 



(11) Pour into sterile Petri dishes and 

 allow the agar to solidify with the 

 cover of the dish removed. 



(12) Dry in the incubator. 

 Sterilization: Sterilize (7) by the fractional 



method. 

 Use: Detection of typhoid group bacilli. 

 The author reported that typhoid colonies 

 were colorless. The medium was trans- 



parent and light green. A great many 

 of the intestinal forms were entirely 

 inhibited. 

 Reference: Padlewsky (1908 p. 543). 



1843. Fildes' Body Fluid Agar 



Constituents : 



1. Extract agar. 



2. Ascitic fluid or other body fiuid. 

 Preparation : 



(1) Collect ascitic fluid in a large glass 

 bottle and allow to stand in the ice 

 chest for a day or two. 



(2) Decant or filter thru glass wool. 



(3) Adjust the reaction to slightly 

 alkaline or neutral to litmus by the 

 addition of alkali or acid. 



(4) Distribute in measured glass stop- 

 pered bottles in 200.0 cc. lots. (The 

 fit of the glass stopper must be 

 perfect.) 



(5) Add chloroform to each bottle until 

 0.5% chloroform has been added. 



(6) .Add a drop of sterile oil to the stop- 

 per and fasten a dust cover tightly 

 over the stopper. 



(7) Place the bottles in the water bath 

 for one hour at 45°C. Shake 

 occasionally. 



(8) When bottles have cooled, remove a 

 sample under aseptic conditions and 

 mix with agar. Incubate the mix- 

 ture at 37°C. for 5 days to test 

 sterility. 



(9) Prepare ordinary lemco (or meat 

 infusion) agar containing 2.5 to 3.0% 

 agar. 



(10) Measure (9) into 200.0 cc. flasks in 

 150.0 cc. lots. 



(11) When ready for use melt in the 

 steamer and treat in one of the 

 following two ways : 



(a) Pour into a sterile distributer 

 maintained at 50°C. by means 

 of a water bath and add 50.0 

 cc. of ascitic fluid (8) for each 

 flask of agar. Mix and tube 

 thru a "hooded" pipette. 



(b) Pour 50.0 cc. of ascitic fluid 

 (8) directly into a flask of 

 melted agar. Mix well, and 

 distribute into tubes or Petri 

 dishes. 



