210 SPECIAL MEDIA 



Animal Tissue Media (Frugoni). 



1. Take a number of sterile test-tubes 16X3 or 4 cm -> P 



with cotton wool, and into each insert a 2 cm. length of stout glass 

 tubing (about i cm. diameter); fill in glycerine (6 per cent.) 

 bouillon to the upper level of the piece of glass tubing. Sterilise 

 in the steamer at 100 C. for twenty minutes on each of three suc- 

 cessive days. 



2. Kill a small rabbit by means of chloroform vapour. 



3. Under strictly aseptic precautions remove the lungs, liver and 

 other solid organs and transfer them to a sterile double glass dish. 



4. With the help of sterile scissors and forceps divide the organs 

 into roughly rectangular blocks 3X1.5X1 cm. 



5. Pour into the dish a sufficient quantity of sterile glycerine 

 solution (6 per cent, in normal saline), cover, and allow to stand 

 for one hour. 



6. Introduce a block of tissue into each tube so that it rests 

 upon the upper end of the piece of glass tubing. (The surface of 

 the tissue will now be kept moist by capillary attraction and 

 condensation) . 



7. Sterilise in the autoclave at 120 C. for thirty minutes. 



8. Cap the tubes and store them in the ice chest for future use. 

 Tissues obtained at postmortems can also be used after pre- 

 liminary sterilisation by boiling or autoclaving. 



Media for the Study of Special Cocci. 



Diplococcus Gonorrhoea. 

 Ascitic Bouillon (Serum Bouillon). 



1. Collect ascitic fluid (pleuritic fluid, hydrocele fluid, etc.), by 

 aspiration directly into sterile flasks, under strictly aseptic pre- 

 cautions. 



2. Mix the serum with twice its bulk of sterile nutrient bouillon 

 (vide page 163). 



3. If considered necessary (on account of the presence of 

 blood, crystals, etc.), filter the serum bouillon through porcelain 

 filter candle. 



4. Tube, and sterilise in the water bath at 56 C. for half an 

 hour on each of five consecutive days. 



5. Incubate at 37 C. for forty-eight hours and eliminate con- 

 taminated tubes. Store the remainder for future use. 



Serum Agar (Heiman). 



i. Prepare nutrient agar (vide page 167), to following formula: 



Agar 2.0 per cent. 



Peptone 1.5 per cent. 



Salt 0.5 per cent. 



Meat extract quantum sufficit. 



