480 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



Pseudomonas trifolii. Other investiga- 

 tors cultivated bacteria found in milk in 

 similar media. 

 Variants : 



(a) Miiller used 1.5% agar instead of 

 2.0%. 



(b) Meier mi.xed equal parts (whey 

 from goat milk (see medium 2277)) 

 and water and dissolved in the mix- 

 ture 1.5% agar, 1.0% Witte's pep- 

 tone (the peptone may be omitted) 

 and 0.5% NaCl. The reaction was 

 adjusted by the addition of KOH 

 until turmeric paper was turned a 

 weak brownish red. 



References: Huss (1907 p. 58), Miiller 

 (1917 p. 390), Meier (1918 p. 435). 



1658. Heller's Peptone Urine Agar 



Constituents : 



1. Urine 1000.0 cc. 



2. Peptone 10.0 g. 



3. NaCl 5.0 g. 



4. Agar (1.5%) 15.0 g. 



Preparation : 



(1) Secure human urine as near average 

 specified gravity as possible. 



(2) Make weakly alkaline with soda. 

 End point indicated by precipitation 

 of salts. 



(3) Filter. 



(4) Add peptone, NaCI and 1.5% agar. 



(5) Boil. 



(6) Filter. 



(7) Tube. 



Sterilization: Sterilize once in streaming 



steam. 

 Use: General culture medium. 

 Variants : 



(a) The author specified that any carbo- 

 hydrate, alcohol, etc., might be 

 added. The color and some in- 

 hibitory substances may be removed 

 by animal charcoal. 



(b) Piorkowski used the following me- 

 dium to differentiate between Bac- 

 terium coli and Bacillus typhi ab- 

 domin. 



(1) Add 100.0 cc. urine and 0.5 g. pep- 

 tone to a flask and plug with cot- 

 ton. Urine should be fresh, clear, 

 bright yellow, and of acid reaction. 

 A urine of specific gravity of 1.012 

 gives the best medium. 



(2) Steam for 15 minutes in a steamer. 



(3) Dissolve 10.0 to 12.0% gelatin or 

 agar in (2). 



(4) Filter, using a hot water funnel or 

 in a steamer. 



(5) Distribute into 10.0 cc. lots and 

 sterilize for 10 to 15 minutes on 

 each of two days using the frac- 

 tional method. 



(c) Piorkowski also gave the following 

 medium for the differentiation of 

 colon and typhoid organisms. He 

 reported that the medium was blue. 

 Bad. coli communi appeared as 

 bluish-grey opaque damp colonies. 

 Typhoid bacillus colonies delicate, 

 transparent and appeared blue. 



(1) Add 100.0 cc. of urine and 0.5 g. 

 peptone to a flask and plug with 

 cotton. A urine with a specific 

 gravity of 1012 gave best results. 



(2) Steam for 15 minutes in a steamer. 



(3) Dissolve 2.0 g. of agar in (2). 



(4) Filter, using a hot water funnel or 

 filter in a steamer. 



(5) Distribute in 10.0 cc. lots. 



(6) To each tube add 8 drops of 

 Bohmer's Hamato.xylyn solution 

 (strength not given). 



(7) Sterilize for 15 minutes on each of 

 two successive days. 



(d) Matzuschita dissolved 2.0% agar, 

 1.0% peptone and 0.5% NaCl in urine. 



References: Heller (1890 p. 893), Pior- 

 kowski (1896 pp. 687, 694), Matzuschita 

 (1901-2 p. 214). 



1659. LoefHer's Malachite Green Nutrose 

 Peptone Agar (Harvey) 



Constituents: 



1. Distilled water 1000.0 cc. 



2. Peptone 20.0 g. 



3. Lactose 50. g, 



4. Glucose 10.0 g. 



5. Nutrose 10.0 g. 



6. Malachite green (2.0% 



chem. pure) lO.O cc. 



7. NaOH (N/1) 15.0 cc. 



Preparation: (1) Dissolve 2, 3, 4, 5, 6 and 7 



in 1. 

 Sterilization: Not specified. 

 Use: Cultivation of colon-typhoid group. 

 Variants: Klimmer gave the following 

 method of preparation: 



(1) Prepare a 0.5% peptone solution. 



(2) Add 9.5 cc. of HCl to (1). 



