52 DIFCOMANUAL 



BACTO 



ENTEROGOGGI GONFIRMATORY BROTH (B302) 



DEHYDRATED 



Bacto-Tryptone 5 g. 



Bacto- Yeast Extract 5 g. 



Bacto-Dextrose 5 g. 



Sodium Azide 0.4 g. 



Sodium Chloride 65 g. 



Bacto-Methylene Blue 0.01 g. 



A discussion of the use of this medium is given above under Bacto-Enterococci 

 Presumptive Broth. 



To rehydrate the medium, dissolve 80.4 grams Bacto-Enterococci Confirma- 

 tory Broth in 1000 ml. of distilled water. Distribute in flasks in 100 ml. quanti- 

 ties and sterilize in the autoclave for 15 minues at 15 pounds pressure (121°G.). 

 Final reaction of the medium will be pH 8.0. When cooled to room temperature 

 and just prior to use, add 65 units of penicillin to each 100 ml. of medium. 

 Enough Enterococci Confirmatory Broth containing penicillin is added to each 

 Enterococci Confirmatory Agar slant to cover approximately one-half the surface 

 of the slant. 



One pound of Bacto-Enterococci Confirmatory Broth will make 5.6 liters of 

 medium. 



BACTO 



BRILLIANT GREEN BILE AGAR (B14) 



DEHYDRATED 



Bacto-Peptone 8.25 g 



Bacto-Lactose 1.9 g 



Bacto-Oxgall 0.00295 g 



Sodium Sulfite 0.205 g 



Ferric Chloride 0.0295 g 



Monopotassium Phosphate 0.0153 g 



Special Agar (Noble) 10.15 g 



Erioglaucine 0.0649 g, 



Bacto-Basic Fuchsin 0.0776 g 



Bacto-Brilliant Green 0.0000295 g 



Bacto-Brilliant Green Bile Agar duplicates the medium described by Noble 

 and Tonneyi for determining the relative density of coliform bacteria in water 

 and sewage. This medium is not recommended for the determination of the abso- 

 lute density of coliform organisms in water samples, but rather as an indication 

 of the degree of contamination of the sample. It is suggested as a selective agar 

 medium for this purpose in Appendix I of "Standard Methods for the Examina- 

 tion of Water and Sewage."^ 



In the enumeration of coliform bacteria using Bacto-Brilliant Green Bile Agar 

 plates should be poured in dilutions which will show not less than ten colonies of 

 coliform organisms per plate and it is, therefore, suggested that several dilutions 

 be plated from each sample. When 10 ml. quantities of water are to be plated 

 an equal quantity of double strength medium should be employed. Inoculated 

 plates are incubated at 37°C. for 17 hours and for not longer than 19 hours. 

 Colonies of coliform bacteria are deep red at the center with a pink halo sharply 



