250 



CULTURE MEDIA FOR CULTIVATION OF MICROORGANISMS 



9.0 NaNOa instead of 5.0 g. KNO3 

 to bouillon. 



(b) Lohnis added about 0.1% NaNOs to 

 bouillon. 



(c) Percival added 3.0 g. NaNO, to 

 bouillon. 



References: Heinemann (1905 p. 131), 

 Sewerin (1909 p. 349), Lohnis (1913 p. 98), 

 Percival (1920 p. 165). 



815. Makgill's Indicator Bouillon 



Constituents : 



1. Bouillon 1000.0 cc. 



2. Neutral red (saturated 

 aqueous solution) 10.0 cc. 



Preparation: (1) Add 1.0% of a saturated 

 solution of neutral red to nutrient 

 bouillon. 



Sterilization: Not specified. 



Use: Detection of Bacillus coli in water 

 analysis. Bacillus coli gave a canary 

 yellow color to the medium. Different 

 authors have added various indicators for 

 other special purposes. These will be 

 indicated under variants. 



Variants : 



(a) Gordon reported that Streptococcus 

 brevis decolorized bouillon contain- 

 ing 2.0 cc. of a 2.0%o watery solution 

 of neutral red per liter while Strepto- 

 coccus longus did not decolorize the 

 medium. 



(b) Heinemann added 10.0% of a 1.0% 

 litmus solution to nutrient bouillon. 



(c) Calandra added 4 drops of a 1.0% 

 aqueous solution of Brilliant cresyl 

 blue to 10.0 cc. bouillon. He re- 

 ported that typhoid bacilli caused no 

 change. B. coli after 24 hours gave 

 a blue colored layer about i cm. high 

 on the upper surface. After 48 hours 

 the blue color persisted in the upper 

 layer. The remainder of the tube 

 was decolorized. 



(d) Calandra added 3 drops of a 1.0% 

 congo red solution to 10.0 cc. of 

 bouillon. He reported that typhoid 

 caused no change in color after 24 

 hours. After 24 hours, B. coli gave 

 a strawberry red color. Both ty- 

 phoid and B. coli were red after 

 48 hours. 



(e) Calandra added 3 drops of a 1.0% 

 Kuhne alkali blue to 10.0 cc. of bouil- 

 lon. He reported that after 48 hours 



typhoid had not changed color. 

 Coli bacilli formed a greenish blue 

 zone of about 1 cm. on the surface of 

 the bouillon. 



(f) Signorelli added 0.5 cc. of a 1.0%, 

 dahlia solution, 10.0 cc. of a 1.0% 

 erythrosin solution or 10.0 cc. of a 

 1.0% safranin solution to 100.0 cc. 

 of bouillon. He studied the adsorp- 

 tion of dye by the cholera vibrio. 

 He reported that the organisms de- 

 veloped slowly. After 24 to 36 hours 

 the colored mass of bacteria settled 

 to the bottom and the medium was 

 decolorized after a week. If the dye 

 was added after the culture had 

 developed the vibrio absorbed the 

 dye. With erythrosin and safranin 

 the bacteria were well colored, but 

 the medium was not decolorized. 



(g) Groenewege added 10.0 drops of 

 methylene blue to 150.0 cc. of bouil- 

 lon. The medium was used to study 

 the reduction of Phytobacter lyco- 

 persicum n. sp. the cause of tomato 

 rot. Partial reduction occurred. 



(h) Botez added 2.0 cc. of a 5.0%o methyl 

 violet solution to 100.0 cc. of bouillon. 

 He reported that B. typhosus grew 

 well. Did not decolorize (reduce) 

 methyl violet. Para typhoid A 

 changed the color to a pale violet. 

 Paratyphoid B completely decolorized 

 the medium after 48 hours. B. coli 

 completely decolorized the medium 

 after 48 hours. B. fecal alcaligenes 

 did not reduce methyl violet. B. 

 Shiga did not grow. Cholera vibrio 

 grew slowly and did not change the 

 color after 10 days. B. pyocyaneus 

 reduced methyl violet, 

 (i) Tanner added 2.0 cc. of 1.0%o neutral 

 red solution to 1000.0 cc. of bouillon 

 containing 1.0% peptone and 0.5% 

 NaCl. 

 (j) Heinemann added 2.0 to 3.0 cc. of a 

 2.0%, solution of Hochst's 120" mala- 

 chite green to bouillon, alkaline to 

 litmus. 

 References: Makgill (1901 p. 431), Gordon 

 (1904 p. 271), Heinemann (1905 p. 128), 

 Calandra (1910 pp. 570, 571), Signorelli 

 (1912 p. 472), Groenewege (1913 p. 26), 

 Botez (1915 p. 489), Tanner (1919 p. 46), 

 Heinemann (1922 p. 35). 



