MILS 451 



blue, by Gram 's method, by Neisser 's method, and by 

 Ziehl-Neelsen 's method. 



Note the presence or absence of altered and unaltered 

 vegetable fibres ; pus cells, blood discs ; cocci in groups 

 or chains, diphtheroid bacilli, Gram negative bacilli or 

 cocci, spores and acid fast bacteria. 



ii. Adapt the final stages of the investigation to 

 the special requirements of each individual sample, 

 thus: 



1 . Members of the Coli=typhoid Group. 



1. Emulsify the deposit from the second centrifu- 

 gal tube (D 2 ) with 10 c.c. sterile bouillon and inocu- 

 late three tubes of bile salt broth as follows : 



To Tube No. i add 2.5 c.c. milk deposit emulsion ( = 25 c.c. orig- 

 inal milk.) 



To Tube No. 2 add i.o c.c. milk deposit emulsion ( = 10 c.c. orig- 

 inal milk.) 



To Tube No. 3 add 0.5 c.c. milk deposit emulsion ( = 5 c.c. orig- 

 inal milk.) 



2. Inoculate tube of bile salt broth No. 4 with i c.c. 

 of the original milk. 



3. Inoculate further tubes of bile salt broth with 

 previously prepared dilutions (see page 445) as follows : 



To tube No. 5 add i.o c.c. from capsule I. 

 To tube No. 6 add o. i c.c. from capsule I. 

 To tube No. 7 add i . o c.c. from capsule II. 

 To tube No. 8 add o. i c.c. from capsule II. 

 To tube No. 9 add i.o c.c. from capsule III. 

 To tube No. 10 add o. i c.c. from capsule III. 

 ' To tube No. n add i.o c.c. from capsule IV. 

 To tube No. 12 add o. i c.c. from capsule IV. 



and incubate anaerobically (in Buchner's tubes) at 

 42 C. for a maximum period of forty-eight hours. 



4. If growth occurs complete the investigation as 

 detailed under the corresponding section of water ex- 

 amination (seepages 428 to 431). 



NOTE. The B. coli communis, derived from the alvine dis- 

 charges of the cow, is almost universally present in large or small 



