ATCC medium: 2155 LYI Giardia medium (filtered)
Available from ATCC as one tube, ready to use upon rehydration. Catalog no. PRA-2155.
LYI Broth (see below).........................88.0 ml
Diamond's Vitamin Mixture 107 (see below)......2.0 ml
Heat-inactivated bovine serum.................10.0 ml
Add Vitamin Mixture and bovine serum aseptically to sterile broth. Dispense medium aseptically in 13-ml aliquots into 16 x 125 mm screw-cap tubes. Cap tightly and store at 4C in the dark. Use within 96 hours.
LYI Broth:
K2HPO4 .........................................1.0 g
KH2PO4 .........................................0.6 g
NaCl...........................................1.0 g
Yeast extract**...............................25.0 g
Neutralized Liver Digest (Oxoid L27)...........5.0 g
L-Cysteine . HCl...............................1.0 g
Dextrose......................................10.0 g
Ascorbic acid..................................0.2 g
Ferric ammonium citrate.......................22.8 mg
Distilled water to...........................880.0 ml
Dissolve the solid ingredients in the order given in 600 ml of the distilled water. Adjust final broth volume to 880 ml with remaining distilled water. Adjust pH to 6.8 with NaOH. Filter-sterilize. Dispense aseptically in 88-ml aliquots into 100-xml screw-cap bottles. Modification of the initial specified formula: 5.0 g/L of the yeast extract has been substituted by 5.0 g/L of Neutralized Liver Digest (Oxoid L27). J. Eukaryot. Microbiol. 42:277-278. 1995.
**NOTE: Some lots of yeast extract may not work. Diamond's Vitamin Mixture (107): Water-soluble B Vitamins (see below)...............500.0 ml
Biotin Solution (see below)........................250.0 ml
Folic Acid Solution (see below)....................250.0 ml
Lipid-soluble Vitamins A, D, and K (see below)...2,500.0 ml
Vitamin E Solution (see below).....................250.0 ml
Sterilize final solution by passage through a 0.22 micron filter. Store the completed, clear mixture at -22C. Thaw and warm to room temperature before use.
If the mixture appears turbid, it should be discarded. Development of turbidity is an indication that an excess of NaOH has been used in the preparation of one of the stock solutions. J. Parasitol. 54: 1047-1056.