4 .T. W. W. STEPHENS — METHODS FOR DETECTING SPOROZOITS 



(e.) Sporozoits are seen in such a fresh preparation in hnndreds as thin, rather 

 glistening, curved rods. Tlie diaphragm should be properly adjusted (fig. .3). 



^=^•''-7"" 



'hT'^ 



Fig. 3. — Sporozoits in the salivary gland. 



Staining of Sporozoits. 



(rt.) Smear off the cover-glass. 



(?>.) Dry as rapidly as possible (over flame). 



(c.) Fix in alcohol for a few minutes — 5 is ample. 



(d.) Stain in Eomanowsky or Leishman, &c. — 10 minutes or longer. 



Compare carefully sporozoits from quartan, simple tertian and malignant 

 tertian cases. Also examine if there is any evidence of c?. Q and indifferent 

 sporozoits. 



Dissection of the mid-gut (stomach). 



N.B — Keep the mosquitos, if caught in huts, for two to three days till the 

 blood is all digested, as it is otherwise almost impossible to see zygotes. 



(«.) Place the mosquito (either a fresh one or the one from which the glands 

 have been extracted) 07i its back in a drop of clean salt solution. With a needle, 

 nick or tear the chitinous body wall on each side as near the " tail " as possible. 

 Place the left needle on the thorax and with the 7-i(jJit needle on the extreme tip 

 make a gentle traction as before. 



(h.) If carefully done, all the viscera will be pulled out, viz., oesophagus and 

 diverticula, mid-gut, Malpighian tubes (5), hind-gut, ovaries (2), &c. When 

 the tension is relieved the mid-gut assumes a flask-shaped saccular appearance. 



(('.) Cut the mid-gut off just below the Malpighian tubes. Also cut through 

 the upper end of the oesophagus to which may be still attached the diverticula--- 

 recognisable by the gas bubbles (CO2) they contain. 



(d.) Remove everything from the slide except the mid-gut (stomach). Add a 

 clean drop of salt solution. Drop on a cover-glass and press gently, so as to 

 flatten out the stomach. Examine with a ]^-inch lens, taking care to adjust the 

 diaphragm properly. 



