INTESTINAL FLAGELLATES IN TISSUE CULTURE 159 



medium and on Boeck's medium of Locke with a few drops of 

 sheep or human serum. Ovomucoid medium is made in the follow- 

 ing way : the white of one hen's egg is shaken up with glass beads. 

 To this is added 100 c.c. of 0.7% sodium chloride solution. This 

 is cooked for half an hour in a glass flask over a hot water bath 

 and kept in constant motion. It is then filtered through cotton with 

 a suction pump and about six cubic centimeters of the filtrate put 

 in small test tubes. These are autoclaved for fifteen minutes at 

 fifteen pounds pressure. 



So far the dififerent species of Giardia have not been grown out- 

 side of their hosts. 



Trichomonas hominis has been grown by many workers, Lynch 

 (1915), Hogue (192 1 6, 1922&), Cleveland (1925), and others. 

 The chief things necessary for the growth of this flagellate seem 

 to be a salt solution with some protein which may be either the 

 white of hen's eggs or the serum of man, pig, or sheep. Both ovo- 

 mucoid and sodium chloride sheep serum water have been success- 

 fully used by the writer. The latter is made in the following 

 way: to a flask containing 100 cubic centimeters of 0.85% sodium 

 chloride solution which has been sterilized in the autoclave add ten 

 to fifteen cubic centimeters of sterile sheep serum water. The 

 serum water is prepared by diluting one part of sheep serum with 

 three parts of distilled water and sterilizing in the autoclave or 

 Arnold sterilizer. 



The pH should be somewhere between 6.8 and 8.4, though 

 Trichomonas lives longest at pH 7.2-7.4 (Hogue, 1922&). The tem- 

 perature of the incubator should be near that of the host, though 

 most forms of protozoa will grow well at a temperature a few de- 

 grees below that of their host. 



The trichomonads which have been used by the writer were 

 obtained (i) from the mouth of man (T. buccalis) by rubbing 

 sterile swabs along the teeth close to the gums and culturing the 

 material obtained in sodium chloride sheep serum water (Hogue, 

 1926) ; (2) from human feces (T. hominis and Pentatrichomonas 

 ardin-dcltcili) , from white rat feces (T. muris), and from chicken 

 feces (T. gallinarum) by putting pieces of feces the size of a small 

 pea in test tubes containing six cubic centimeters of sodium 

 chloride sheep serum water; (3) from the vagina (T. vaginalis) 

 using sterile swabs and culturing in sodium chloride sheep serum 

 water. 



