ATCC medium: 1967 Soil freshwater cereal grass medium
ATCC Medium 802 (See below)......750.0 ml
ATCC Medium 1900 (see below).....250.0 ml
Prepare sterile batches of ATCC media 802 and 1900 separately. Aseptically combine cooled solutions in the amounts specified above.
ATCC Medium 802:
Solution 1 Rye grass cerophyll:
Cerophyll*.........................2.5 g
Distilled water....................1.0 L
Add cerophyll to distilled water and boil for 5 minutes. Add 100 ml distilled water to compensate for evaporation. Filter through Whatman #1 filter paper and add 0.5 g Na2HPO4. Autoclave for 15 minutes at 121C.
Solution 2 Agar for Klebsiella pheumoniae:
Agar..............................20.0 g
Yeast extract......................4.0 g
Glucose............................0.16 g
Distilled water..................800.0 ml
Dispense in 5.0 ml amounts. Autoclave for 25 minutes at 121C. Slant. Bacterium, grown on solution 2, is added to Solution 1 and incubated at 30C for 24 hours prior to inoculation with Paramecium. *Can be obtained from Ward's Natural Science Establishment, Inc. P.O. Box 92912, Rochester, NY 14692. Dairy Goat Nutrition P.O. Box 22363, Kansas City, MO 64113 distributes Grass Media Culture which is equivalent. Sigma Chemical Company, P.O. Box 14508, St. Louis, MO 63178, sells Cereal Leaf Product no. C-7141 which is similar to Cerophyll.
ATCC Medium 1900:
Soil Extract Stock (see below)...625.0 ml
Solution 1 (see below)...........125.0 ml
Solution 2 (see below)...........125.0 ml
Solution 3 (see below)...........125.0 ml
Mix all components and filter-sterilize.
Soil Extract Stock Air-dried sieved soil............333.3 g
Distilled water..................666.7 ml
Adjust pH to 8.0 with N NaOH or HCl. Autoclave for 30 minutes. Allow soil to settle and decant. Pre-filter through cheese cloth to remove large particles.
Solution 1:
K2HPO4 .............................1.0 g
Distilled water....................1.0 L
Solution 2:
MgSO4 . 7H2O .......................1.0 g
Distilled water....................1.0 L
Solution 3:
KNO3 ..............................10.0 g
Distilled water....................1.0 L