ZOOLOGY AND BOTANY, MICROSCOPY, ETC. 249 



(4) Staining and Injecting:. 



New Modification of the Romanowsky-Ruge Method for Staining 1 

 Blood-Spores.* Berestneff recommends the following. Stain No. 1 : 

 • 5 p.c. watery solution methylen-blue (med. puriss.). Stain No. 2 : 1 p.c. 

 watery solution methylen-blue and ■ 3 p.c. crystalline soda, heated for 

 three hours in a water bath and then filtered. Stain No. 3:0*5 p.c. 

 watery solution eosin (extra B.A.). Four parts of No. 1 are mixed with 

 one part of No. 2, and to 5 c.cm. of this 2 "25 c.cm. of No. 3 are added. 

 The preparation is fixed in absolute alcohol, and then stained for 10 to 

 30 minutes (crescents require at least 35 minutes). The preparation is 

 then dried with filter-paper, or quickly washed with water, differentiated 

 in a mixture of 100 parts alcohol and 2 parts 5 p.c. acetic acid for a 

 few seconds, washed quickly in water and dried. 



Demonstrating Presence of Cilia in Bacteria.f — D. Ellis used 

 ordinary agar, " spirillum agar," and peptone-beef broth, and his method 

 was to keep on continually transferring the organism to a fresh medium 

 as soon as growth was perceptible. He was successful in demonstrating 

 cilia in all these species. The following staining method was employed : — ■ 

 Three small drops of water were placed on an absolutely clean slide. A 

 portion of the material was then, with a platinum loop, mixed with the 

 first drop. A loopful of this drop was then mixed with the second, and, 

 lastly, a loopful of the second with the third. From the third drop the 

 cover-glass preparations were made. The smears were then fixed by 

 being kept at 37° C. for 4 minutes, then mordanted for 3^-7 minutes 

 with — 



10 c.cm. of a 20 p.c. sol. of Tannin, 

 8 c.cm. of a cold sat. sol. FeS0 4 , 

 1 c.cm. of a sat. sol. of Fuchsin in Abs. Ale. ; 



and, lastly, stained for 5 minutes with — 



1 grm. Saure violett (Griibler & Co. 6 B), 

 75 c.cm. Absolute Alcohol, 

 75 c.cm. water. 



Resistance of Tubercle and other Acid-fast Bacilli to Decolorising 

 Agents.* — C. A. Coles submitted the bacilli of tubercle, smegma, 

 Timothy grass, grass bacillus ii. and mist bacillus to various decolorising 

 agents, after staining with Ziehl-Nielsen for seven minutes. The most 

 important results are that tubercle bacilli can resist 25 p.c. sulphuric 

 acid for 72 hours, while pseudo-tubercle bacilli are decolorised in 

 16 hours or less. Tubercle bacilli resist Pappenheim's solution [1 part 

 corallin (rosolic acid) in 100 parts of absolute alcohol, to which methylen- 

 blue is added to saturation ; this mixture is further treated with 20 parts 

 of glycerin] for 52 hours, while pseudo-tubercle bacilli are decolorised 

 at the end of four hours. 



The author suggests a modification of Pappenheim's solution, finding 



* Centralbl. Bakt, V Abt. Ref., xxxiv. (1904) p. 296. 



t See ante, p. 232. 



X Repr. from Journ. State Med., Feb. and March, 1904, 20jpp. 



April 20th, 1 90 4 3 



