Euglenidae 63 



CULTURE OF SOME FLAGELLATES AND CILIATES 

 Paul Brandwein, Washington Square College, New York University 



Euglena. To 100 cc. of a modified Klebs' solution (Solution B)* in 

 a white glass battery jar add 40 rice grains (boiled 5-10 minutes) and 

 900 cc. of distilled water. The foregoing medium is allowed to stand for 

 about five days. The jar is then placed in indirect sunlight (the direct 

 rays of the sun should not strike this culture for more than an hour a 

 day), and inoculated with Euglenae three times (10 cc. of a dense 

 Euglena culture) at three day intervals. If an old Euglena culture is 

 available the organisms may be found encysted on the sides of the vessel 

 and it is of great advantage to inoculate the cysts along with the free 

 Euglenae. Starting ten days after the initial inoculation, growth may be 

 accelerated by adding (three times, at weekly intervals), 25 cc. of Solu- 

 tion B and 10 mg. of the tryptophane powder. The further addition of 

 5 grains of boiled rice each month will serve thereafter to maintain the 

 culture. Large ciliates and rotifers are detrimental. 



Another technique, applicable to several Protozoa involves the use of 

 an egg yolk-distilled water medium. 



Chilomonas. A thin smooth paste is prepared by grinding 0.5 gm. of 

 the boiled yolk of a fresh hen's egg with a small amount of distilled water. 

 This is added to 500 cc. of distilled water and the mixture, after standing 

 two days, is inoculated with the original Chilomonas culture. If such a 

 culture is not available, spontaneous inoculation will occur if the culture 

 jar is left uncovered, since cysts of Chilomonas seem to be omnipresent. 



Paramecium, Colpidium, Colpoda, Euplotes. These ciliates have done 

 well on the egg yolk medium when Chilomonas is provided as prey. Start 

 a Chilomonas culture as previously directed and inoculate with 10 cc. 

 of a culture of the desired ciliate, three times, on the 4th, 6th, and 8th 

 day after starting. Dense maximum growth has usually been obtained 

 in two weeks and subculturing has been necessary about every month. 



Didinium. The organism will thrive exceedingly well when introduced 

 into one of the Paramecium cultures described above. As the Para- 

 mecium diminishes in one culture a fresh one should be available for 

 inoculation with the Didinium. In fact it is advisable to keep 

 Paramecium cultures in a separate room ; otherwise it is difficult to avoid 

 contamination with Didinium. 



Vorticella. A modification of the method found useful for Paramecium 

 is necessary here. The medium of % gram of mashed hard-boiled egg 

 yolk in 750 cc. of distilled water is permitted to stand for two days; it 

 is then filtered through cotton, 100 cc. of the filtrate are added to a 



♦Modified Klebs' Solution (Solution B): KXO :J .25 gm., MgS0 4 .25 gm., KH 2 P0 4 

 .25 gm., Ca(N0 3 ) 2 1 gm., Bacto-Tryptophane Broth (powder) .010 gm. (Digestive 

 Ferments Co.), distilled water to make 1000 cc. 



