An obligative anaerobic fermentation Sarcina. 



Proceedings of the Section of Sciences, Kon. Akademie van Wetenschappen, Amster- 

 dam, Vol. VII, 1905, p. 580 585. -- Verscheen onder den titel Een obligaat anaerobe 

 gistingssarcine in Verslagen Kon. Akademie van Wetenschappen, Wis-en Natuurk. Af- 

 deeling, Amsterdam, Deel XIII, 1905, biz. 608 614; en onder den titel Une sarcine 

 de fermentation anaerobic obligatoire in Archives Neerlandaises des Sciences Exactes 

 et Naturelles, Haarlem, Serie II, Tome XI, 1906, p. 199 205. 



The following simple but yet delicate experiment gives rise to a vigorous fer- 

 mentation, caused by a sarcine, wherein microscopically no other microbes 

 are perceptible and which, when rightly performed, can produce a real pure culture 

 of this fermentation organism. The simplicity of the experiment is the result of 

 many previous investigations, partly made conjointly with Dr. N. Goslings, 

 which have gradually rendered clear the conditions of life of the examined microbe. 



Bouillon with 3 to 10% glucose, or malt wort, is acidified with phosphoric acid 

 to an acidity of 8 cc. normal per 100 cc. of culture liquid and introduced into a bottle, 

 which is quite filled with it and fitted with a tube to remove the gas. The infection is 

 done with an ample quantity A ) of garden soil, from which the heaviest and roughest 

 portion has been removed, but in which so much solid substance is left behind that 

 in the nutrient liquid it forms a muddy deposit from 5 to 7 or more millimeters thick. 

 The culture is effected in a thermostat at 37 C. After 12 hours already the liquid 

 is in a strong fermentation, which lasts from 24 to 36 hours, and whereby the surface 

 is covered with a rough scum, produced by gas bubbles mounting up from the depth. 

 Whilst the liquid itself remains wholly free from microbes, the microscopical image 

 of the deposit shows a luxuriant, pure or almost pure culture of a sarcine, of which 

 the elementary cells measure for the greater part about 3.5 (a, so that the species 

 belongs to the largest forms known, and the multicellular sarcine-packages are easily 

 visible to the naked eye. The cells are colorless and transparent and the packages 

 present irregular sides. Here and there, but much less generally, a brownish intrans- 

 parent form is seen, with more regularly cubical packages of which the cells measure 

 2 to 2,5 jn. 



The scum floating on the fermenting fluid consists of slime in which the evolved 

 gas remains for a time imprisoned. This slime is produced by the outer side of the 

 sarcine cells, whose walls for the rest consist of cellulose, which becomes violet-blue 

 by zincchloride and jodine. This reaction was discovered in 1865 in the stomacal 

 sarcine by Suringar 2 ), who on this account argued the vegetal nature of this 



*) With little soil for infection, the experiment becomes doubtful. 

 2 ) W. F. R. Suringar, De sarcine (Sarcina ventriculi Good sir), pag. 7, 

 Leeuwarden 1865. Here very good figures are to be found. 



