CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



729 



of acid reaction. A urine of specific 



gravity of 1.012 gives the best medium. 

 Variants: Piorkowski prepared similar 



media as follows: 



(a) He reported that on the following me- 

 dium Bad. coli commune colonies 

 after 20 hours, under the microscope, 

 were round, yellow fine grained and 

 edges were well defined. Bad. typhi 

 ahd. gave thread like colonies. Do 

 not incubate at a temperature lower 

 than 22°C. 



(1) Allow normal urine to stand for 

 2 days. Specific weight of the urine 

 to be 1.020. During this time the 

 reaction has become alkaline. 



(2) Add 0.5% peptone and 3.3% gelatin 

 to (1). 



(3) Boil for an hour in a water bath. 



(4) Filter at once without the use of 

 heat. 



(5) Distribute into test tubes and plug 

 with cotton. 



(6) Sterilize in the steamer for 15 min- 

 utes at 100°C. and on the following 

 day sterilize at 100°C. for 10 

 minutes. 



(7) Incubate at 22°C. 



(b) On the following medium typhoid 

 colonies were small and colorless, 

 with 4 to 6 turft like runners. Bad. 

 coli colonies were large and yellow 

 without the runners. 



(1) Dissolve 0.5% peptone and 3.3% 

 gelatin in alkaline urine having a 

 specific gravity of 1.020. 



(2) Allow to stand at 22° for 16 to 

 20 hours. 



(3) Sterilization not specified. 

 References: Piorkowski (1896 p. 687), (1899 



p. 145), (1916-17 p. 259). 



2279. Heller's Urine Peptone Gelatin 



Same as medium 745 but solidified by the 

 addition of 10.0% gelatin. 



2280. Schultz's Peptone Infusion Gelatin 



Constituents: 



1. Water 1300.0 cc. 



2. Meat 500.0 g. 



3. Peptone (siccum). . . 10.0 g. 



4. NaCl 5.0 g. 



5. Gelatin 50.0 to 100.0 g. 



Preparation : 



(1) Place 500.0 g. of the best quality 

 meat, without fat or tendons, in a 

 glass container fitted with a lid. 



(2) Pour 1300.0 cc. distilled water over 

 the meat. 



(3) Store in a cool place until the next 

 day. 



(4) Filter thru four thicknesses of 

 cloth and press the remaining meat 

 to obtain as much fluid as possible. 



(5) Pour the filtrate into a kettle, add 

 10.0 g. peptone (siccum) 5.0 g. NaCl 

 and the whites of two eggs, beaten 

 up in two or three volumes of water. 



(6) Boil over a gas flame for 15 minutes. 



(7) Adjust to faint alkalinity, using 

 phenolphthalein as an indicator, and 

 the end point being a faint red color. 



(8) Pour into an iron kettle, add 100.0 cc. 

 distilled water, boil strongly for 

 5 minutes and filter. 



(9) Dissolve 50.0 to 100.0 g. of gelatin 

 in (8). 



(10) Add 200.0 cc. distilled water to (9). 



(11) Cool to 40°C. and add the whites of 

 two eggs beaten up in 2 or 3 volumes 

 of water. 



(12) Boil strongly for 10 to 15 minutes. 



(13) Filter thru four layers of cloth and 

 then thru a hot water funnel. 



Sterilization: Sterilize for 30 minutes on 

 each of 3 successive days in a steamer. 



Use: General culture medium. 



Variants : 



(a) Acosta and Grande (1892) (Sentinon 

 1893). 



(1) Add to 1000.0 of tendon free lean 

 meat a double weight of water. 



(2) Boil (time not specified). 



(3) Skim and strain. 



(4) Place once more on the fire and 

 and 0.5% peptone and 0.25% NaCl 

 and add water to bring up to the 

 original volume. 



(5) Add 16.0 to 18.0% gelatin. 



(6) Pour the solution in a clay or glass 

 container that is twice as high as 

 wide. 



(7) Autoclave at 105° at 0.5 atmos- 

 phere pressure for 15 minutes. 



(8) Release the pressure and allow the 

 mi.xture to stand for 24 hours. 



