ZOOLOGY AND BOTANY, MICROSCOPY, ETC. 3()5 



middle, others joining to form filaments, and others resembling spirilla. 

 They are most frequent in the parts in condition of grey hepatization. 

 The author mentions that in some of the preparations a Gram-positive 

 streptococcus was present. Cultivations were difficult ; in only one tube 

 was a pure growth obtained, the rest being contaminated with the 

 streptococcus. The medium used was agar mixed with bouillon Martin 

 and calf's blood. There is no record of animal experiments. 



Enrichment and Staining Methods for Tubercle bacilli.* — W. 

 Frei found that the best enrichment method for demonstrating tubercle 

 bacilli in sputum was that of Hammerl (dissolving the sputum in a 

 mixture of ammonia and caustic potash, shaking up with acetone and 

 centrifuging). Other less effective methods were (1) the antiformiu 

 (sputum dissolved in 20 p.c. antiformin, centrifuged) ; (2) antiformin- 

 ligroin (sputum dissolved in 20 p.c. antiformin, shaken up with ligroin ; 

 sedimented) ; (3) antiformiu-chloroform (sputum dissolved with heat in 

 50 p.c. antiformin, shaken up with chloroform-alcohol, centrifuged). 

 For staining, besides the Ziehl-Neelsen, Much's modification of Gram's 

 method, and Herman's method were used. Tlie Much-Gram procedure 

 adopted was as follows : Stain with solution of 10 c.cm. of saturated 

 alcoholic solution of methyl- violet B.N. in 100 c.cm. of 2 p.c. carbolic 

 acid water for 24 to 4.S hours in an incubator at 37°, or by boiling over 

 the flame. Then treat for 1 to 5 minutes with iodo-potassic iodide solu- 

 tion, followed by 1 minute in 5 p.c. nitric acid, 10 seconds in 3 p.c. 

 hydrochloric acid ; aceton-alcohol aa, contrast-stain with dilute carbol- 

 fuchsin. The tubercle bacilli are seen as rows of 4-6 granules. 



Herman's staining method : a freshly made mixture of 3 parts of 

 1 p.c. ammonium carbonate in distilled water and 1 part of 3 p.c. 

 crystal- violet solution in 95 p.c. alcohol, is poured over the smear and 

 heated in the flame until steam is given off : the preparation is allowed 

 to stain for a minute, after which it is treated for several seconds with 

 10 p.c. nitric acid and then with 95 p.c. alcohol. This last procedure 

 is repeated until the preparation assumes a pale blue hue. A 1 p.c. 

 aqueous or alcoholic solution of eosin is used as contrast-stain. Other 

 methods of counter-staining may be adopted. 



Staining Blood-plates in Sections of Organs.! — L. Le Sourd and 

 P. Pagniez describe a method for staining the blood-plates in sections. 

 The pieces are fixed in Dominici's fluid for 12 to 15 hours, and then em- 

 bedded in paraffin. The sections are stained twice in Giemsa's solution. 

 The first time for 12 to 15 hours, Giemsa 5 drops to 15 c.cm. of distilled 

 water. The second time for 4 to 5 hours in Giemsa 15 drops, distilled 

 water 15 c.cm. On removal the sections are at once treated successively 

 with the following mixtures of acetone and xylol : 1. Acetone 18 drops, 

 xylol 2 drops. 2. Acetone 14 drops, xylol 6 drops. 3. Acetone 6 drops, 

 xylol 14 drops. These mixtures are dropped over the section, and when 

 there is good differentiation the preparations are treated with pure xylol 

 and mounted on balsam. The staining is not very permanent. 



-" Centralbl. Bakt. Ite Abt. Orig., Ixi. (1911) pp. 411-16. 

 t C.R. Soc. Biol. Paris, Ixxi. (1911) pp. 308-10. ; 



