. A laboratory guide in bacteriology, for the use of students, teachers, and practitioners. loopful of serum. o) Prepare a thin film, and dry in the air. b) When dry, cover with a 5 to 10 per cent solutionof tannic acid for 10 to 20 seconds. c) Wash in water and dry with blotting paper. d) Cover with carbol gentian violet or anilln gentianviolet for one-half to one minute. Carbol gentianviolet is prepared by mixing one part saturated alco-holic solution of Griiblers gentian violet with 4 partsof an aqueous 5 per cent phenol solution. e) Wash in water. /) Stain with Grams iodin solution for one
. A laboratory guide in bacteriology, for the use of students, teachers, and practitioners. loopful of serum. o) Prepare a thin film, and dry in the air. b) When dry, cover with a 5 to 10 per cent solutionof tannic acid for 10 to 20 seconds. c) Wash in water and dry with blotting paper. d) Cover with carbol gentian violet or anilln gentianviolet for one-half to one minute. Carbol gentianviolet is prepared by mixing one part saturated alco-holic solution of Griiblers gentian violet with 4 partsof an aqueous 5 per cent phenol solution. e) Wash in water. /) Stain with Grams iodin solution for one-half toone minute. g) Decolorize in alcohol. IMPORTANT PATHOGENIC BACTERIA III h) Stain with a saturated alcoholic (60 per cent)solution of Griibers eosin. i) Wash in water and blot. j) Clear and mount in balsam. Whichever method is appUed, the capsule shouldappear as a lightly stained zone with a well-definedoutUne around the deeply stained cell. Capsules arerarely demonstrable unless the organisms are culti-vated in media rich in proteins, or are mixed withserum previous to Fig. 32Mouse-Holder Special study.—^Inoculation of a mouse with 1. Fasten the mouse in the holder (Fig. 32). 2. Shave a place on the back above the tail. 3. Wash with a solution of mercuric chlorid(1:1,000), followed by alcohol. 4. Inject of a milk culture of Str. pneu-moniae. 5. When dead, perform an autopsy, and study thelesions in the usual manner. 6. Make cultures in milk and on slant agar fromthe hearts blood or the spleen. 7. Make a capsule stain from the hearts blood,spleen, or other organs. 112 LABORATORY GUIDE IN BACTERIOLOGY SECTION 3THE GROUP OF COLON-TYPHOID BACILLI This chapter is devoted to the study of the groupof intestinal organisms. This collective group mayconveniently be subdivided into three subgroups: Subgroup I: the colon group.—^This group includesdifferent varieties of Bacillus coli and B. aerogenes. Subgroup 2: the B. enteritidis group.—^
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