44 RESEARCH IN PROTOZOOLOGY 



be studied also from normally fed hosts, as alterations of cyto- 

 plasmic inclusions, body form and size proportions may result 

 from cellulose feeding or starvation. 



For permanent preparations smears made from the intestine of 

 the living termite are most satisfactory. These smears can be made 

 without dilution or with a small amount of suitable fluid. In the 

 field, smears may be made on circular cover glasses and these 

 stored in vials of eighty-five per cent alcohol with paper rings be- 

 tween them. In order to facilitate removal of the cover slips, a loose 

 wad of cotton should be placed in the bottom of the vial and be- 

 tween groups of the covers. These vials should not be corked but 

 can be stored in glass covered jars of alcohol. The intestinal canal 

 of the termite may be pulled out and fixed entire, for later sec- 

 tioning or smearing, but it is much more difficult to make satis- 

 factory preparations from such material than from smears. Smears 

 may also be made on slides and these stored in jars of alcohol, but 

 in the field slides are more difficult to handle. If material is to be 

 sent to a laboratory for study, smears should be made and entire 

 intestines fixed in the field, in addition to the transportation of 

 living material if this is possible. During transportation some 

 termites may lose part or all of their fauma, though this has been 

 true of only a small proportion of shipments. 



The many methods of fixation and staining which have been 

 used do not differ from general methods in use by protozoologists, 

 so that it is unnecessary to recount them all in detail. It is, however, 

 worth while to point out those methods which have been used 

 with most success. In fixation, best general results have been re- 

 ported from the use of Schaudinn's fluid with from four to five 

 per cent acetic acid and Bouin's fluid, either cold or heated to 

 about 55° C. After use of these reagents, the parabasal bodies 

 usually do not stain well. To demonstrate these, strong Flemming's 

 without acetic, Da Fano's fixative and Champy's fluid have been 

 used. Osmic vapor is an excellent fixative for certain purposes, 

 especially demonstration of flagella, parabasal bodies, blepharo- 

 plasts and yarious granules in the cytoplasm. The smears are ex- 

 posed for a short time, less than a minute, to the vapor of one 

 per cent osmic acid, as recommended by Grasse (1926), and are 

 then stained in iron hematoxylin. 



For staining, Heidenhain's hematoxylin has been most widely 

 used, and is unsurpassed for study of most details of cytological 



