vin CULTIVATION OF VIBRIOS 37 



(5) From the alcohol transfer it directly to filtered 

 aniline gentian-violet and allow it to stain for two to ten 

 minutes. 



(c) Rinse the section in distilled water. 



(d) Now place it in Gram's iodine solution till it is 

 black (about two minutes). 



(e) Transfer it to absolute alcohol for a quarter to half 

 a minute, and thence to 



(/) Acid alcohol (alcohol with 3 per cent hydrochloric 

 acid) for not more than ten seconds. 



(g) Again wash it in alcohol until the section appears 

 colourless, or if previously stained with picrocarmine, until 

 the section again becomes red. 



(ti) Clear it in xylol or turpentine and creasote. 

 (i) Transfer it to a slide with a cigarette-paper. 

 (Jc) Mount it in Canada balsam. 



Examine with low and high powers and ^ i n - ^ 

 immersion : the bacilli appear dark blue on a 

 yellowish or red background. 



Cholera 



Inoculate 



(1) a gelatine tube (stab), an agar-agar tube (streak), 

 and a broth tube with Vibrio cholerse Asiaticae ; 



(2) a gelatine tube (stab), an agar-agar tube (streak), 

 and a broth tube with Vibrio Finkler-Prior ; 



(3) a gelatine tube (stab), an agar-agar tube (streak), 

 and a broth tube with Vibrio Metchnicovi. 



