SPIRILLUM CHOLERA ASIATICS. 417 



observed that the comma bacilli usually multiply rapidly 

 on moist linen or moist earth, and that it is not till 

 after 2 or 3 days that other forms of bacteria have 

 gained the upper hand. Hence shirts and bed-clothes, 

 even when they have been kept for some time, are as a 

 rule still suitable for the discovery of the comma bacilli. 

 The microscopical investigation was made by spreading Microscopical 

 out on cover glasses a drop of the dejecta, or, better, one investi & ation - 

 of the small mucous flakes which are usually present in 

 large numbers in the turbid fluid, and which contain 

 the largest numbers of the comma bacilli ; these cover 

 glasses were then dried, heated, and stained for 1 to 5 

 minutes with warm methylene blue or fuchsirie solutions. 

 It is often difficult to recognise the comma bacilli with Modification 

 certainty among the large number of other bacteria ; S 

 according to Schottelius it is of advantage in these 

 cases, after mixing the dejecta with double the quantity 

 of faintly alkaline meat infusion, to let it stand in an 

 open vessel for 12 hours in a warm place (30 to 40 C.). 

 Under these circumstances the comma bacilli multiply 

 chiefly at the surface of the fluid, and by taking a speci- 

 men from the surface, preparations are obtained which 

 contain almost exclusively comma bacilli in such numbers 

 as are only otherwise found in the typical soupy dejecta. 

 It is more difficult to demonstrate the comma bacilli in Demonstra- 

 sections of the intestinal mucous membrane. In cases tion of comma 



bacilli in sec- 



which follow a very acute course the bacilli scarcely tions of the 

 seem to penetrate into the mucous membrane at all ; in mucous* 1 

 old cases the bacilli which have passed into the mem- membrane - 

 brane have long since died, and only other bacteria 

 which entered afterwards are found. The comma bacilli 

 are best demonstrated in sections of intestinal mucous 

 membrane which is in the state of patchy redness de- 

 scribed above; under these circumstances they can be 

 almost always found in sections through the red border 

 of the follicles. They are best stained with alkaline 

 methylene blue ; a suitable method of double staining 

 has not as yet been found, and this is so much the 

 more to be regretted as it is by no means easy to see 

 the comma bacilli in the midst of the cells and nuclei 



27 



