488 METHODS AND FORMULAS DS 23.217 



23.217 Gutstein 1926 23684, 93 :393 



REAGENTS REQUIRED; A. sat. sol. ammonium sulfate; B. 5% tannin; C. 1% malachite 



green 

 method: [air-dried smear] -^ A, 5 mins. —> rinse —* B, 2 mins. — » thorough wash -^ C, 



5-60 sees. — » wash -^ dry -^ balsam 



23.217 Hiss 1905 11189, 6:317 



REAGENTS REQUIRED: A. 0.1% Crystal violet; B. 20% copper sulfate 

 method: [heat-fixed smears] — > A, on slide, warmed to steaming, 3^^ to 1 min. — » B, till 



no more color comes away — * blot — » dry 

 result: capsule blue. 

 note: Anthony 1931 (19938, 73:319) substitutes 1% crystal violet for A above. Tyler 



test. 1946 Conn and Darrow cit. Park and Williams 1933 (Conn and Darrow 1946, 



111A2-18) adds 0.25% acetic acid to A above. 



23.217 Huntoon 1917 11056,2:241 



reagents required: A. 3% nutrose; B. water 100, phenol 2, lactic acid 0.5, acetic acid 



0.01, magenta 0.1, DS 11.43 Ziehl 1882 1 

 method: [smear, of culture diluted with A, air-dried]^ B, 30 sees.—* water, wash—* 



dry 

 note: Solution A must be steam-stertlized (not autoclaved) and preservative added if 



required to keep. 



23.217 Johne test. Kahlden and Laurent 1896 Kahlden and Laurent 1896, 100 



reagents required: A. sat. aq. sol. crystal violet; B. 1% acetic acid 

 method: [smears] —> A, 25 to 30 sees. -^ B, 5-10 sees. — » dry 



23.217 Kahlden and Laurent 1896 Kalilden and Laurent 1896, 104 



reagents required: A. water 60, crystal violet 0.5, acetic acid SO; B. 1% acetic acid 

 method: [smears] —* A, 2i hrs. 37°C. -^ B, wash -^ balsam, via usual reagents 



23.217 Klett test. 1928 Schmorl Schmorl 1928, 359 



reagents required: A. 1% methylene blue in 10% ale; B. 1% magenta in 10% ale. 7 

 method: [heat-fixed smears] — » A, warmed to steaming, 3-5 mins. -^ wash -^ B, 5 sees. 

 — » wash and dry 



23.217 La wson 1940 11284,25:435 



reagents required: A. 5% phosphomolybdic acid; B. DS 13.12 Wright 1910 (working 



solution) 60, glycerol 30 

 method: [air-dried smears] — > A, on slide, 30 sees. —* wash —>■ wash, methanol — > 10-20 

 drops water, 10-20 mins. — > rinse — » dry 



23.217 Muir 1916 11431, 20:257 



reagents required: A. DS 12.15 gentian violet; B. ADS 12.2 Gram 1884; C. water 100, 

 mercuric chloride 2, potassium alum 1.3, tannic acid 7; D. sat. sol. (circ. 35%) eosin; 

 sat. sol. {circ. 3.5%) potassium alum" 

 method: [mercuric-chloride-fixed, and acetone-washed, smears]^ A, on slide, 2 mins. 

 — > water, rinse -^ B, on slide, 1 min. -^ water, wash -^ methanol 1 min. — ♦ clove oil, 

 heated to fuming, 1 min. -^ methanol, wash —^ water, wash — * C, 5 mins. — > water, 

 rinse -^ D, 30 sees. — * water, wash —* E, 1 min. — > water, rinse —y dry 



23.217 Muir test. 1920 Stitt Stitt 1920, 55 



reagents required: A. DS 11.43 Ziehl 1890; B. water 100, mercuric chloride 1.5, tan- 

 nic acid 4, potassium alum 3; C. 1% methylene blue 

 method: [heat-fixed smears]-^ A, heated to steaming, 30 sees. -^• 95% ale, rinse—* 

 water, wash — * B, 5-10 sees. —* water, wash — » 95 % ale, 1 min. -^ water, wash — » 

 C, 1 min. —>■ dry 



23.217 Raebiger test. 1904 Besson Besson 1904, 166 



reagents required: 40% formaldehyde 100, crystal violet 15 

 preparation: Heat to boiling. Cool. Filter. 

 method: [air-dried smears] — » A, 1 min. —* wash — > dry 



