TESTICULAR INFUSION AGAR 351 



ence in handling gonococcus cultures. I am pleased to thank 

 Miss Vera Bennett who has prepared much of the culture media 

 for me and Miss Lettie Watkins who has assisted in keeping 

 the stock cultures alive and planting the experimental media. 



SUMMARY 



The formula now followed in preparing testicular infusion 

 agar for the growth of the gonococcus in preparing suitable 

 vaccines. 



1. Mix 500 grams ground beef testicle from which the tunica 

 vaginalis has first been stripped, with 1000 cc. distilled water. 



2. Soak overnight at room temperature. 



3. Heat to 50°C. Keep warm for one hour by placing in the 

 incubator at 37°C. 



4. Boil, strain, and restore to 1000 cc. with distilled water. 

 If in excess do not reduce by boiling since overheating is in- 

 jurious. 



5. Add 2 per cent peptone (Witte's or Difco), 3 per cent agar 

 chopped fine, 0.5 per cent glucose, 0.3 per cent NaH2P04. 



6. Soak at least one hour to soften the agar. 



7. Melt in the autoclave at 10 pounds pressure for 30 minutes. 



8. Titrate with phenolphthalein and add N/1 NaOH sufficient 

 to neutrahze if no unsaturated compounds were present. 



9. Check the titre by repetition. 5 cc. should require from 

 1.0 to 2.0 cc. N/20 NaOH. to display color hot. 



10. Tube and sterilize in the autoclave at 10 pounds for 30 

 minutes. 



11. Slant or pour into plates. 



This medium may be melted for plating, etc., but the less 

 heating the better. Filtration for the purpose of removing the 

 distinct turbidity of the medium also seems to be a disadvantage. 



