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DSMZ

DSMZ 585a . BCYE alpha agar

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Equipment needed: Autoclave Filter
Final pH: 6.9
Select a strain (optional):

ml
Compound Amount Unit
Solution A 490 ml
Solution B 490 ml
Solution C 10 ml
Solution D 10 ml
1 Prepare OXOID Legionella CYE-Agar base or solution A.
2 Cool down to 50 °C.
3 Add solution C (Cysteine-HCl) and solution D (ferric pyrophosphate).
4 Adjust pH to 6.9 with sterile 1.0 N KOH or HCl.
5 The pH of the medium is critical.
6 Finally, add solution B. Swirl medium in flask during dispensing to petri dishes or tubes in order to keep charcoal suspended.

OXOID Legionella CYE-Agar base #

1000
Compound Amount Unit
OXOID Legionella CYE-Agar base
(with Legionella BCYE supplement) 		1000 ml
1 Dehydrated media preparations are available from media distributors.
2 We use OXOID Legionella CYE-Agar base with Legionella BCYE supplement.
3 Note: Alternatively use CYE-ACES AGAR

Solution A #

490
Compound Amount Unit
ACES
(N-2-acetamido-2-aminoethane-sulfonic acid) 		10 g
Yeast extract
(Difco 0127) 		10 g
Activated charcoal
(Sigma C 5510) 		2 g
Na2 alpha-ketoglutarate 1 g
Distilled water 490 ml
1 Dissolve ACES and bring to pH 6.9 by slowly adding 1 N KOH. Add the other components and dissolve by boiling. Autoclave at 121°C for 15 minutes. Cool to 50 - 55°C.

Solution B #

490
Compound Amount Unit
Agar 15 g
Distilled water 490 ml
1 Autoclave at 121°C and cool to 50 - 55°C.

Solution C #

10
Compound Amount Unit
L-Cysteine HCl x H2O 0.4 g
Distilled water 10.0 ml
1 Prepare a fresh solution and filter-sterilize it.

Solution D #

10
Compound Amount Unit
Fe4(PO4)2 0.25 g
Distilled water 10.00 ml
1 Prepare a fresh solution by heating to 50 - 55°C in a water-bath. Filter-sterilize solution separately.
2 Please note: Soluble ferric pyrophosphate must be kept dry and in the dark. Do not use if chemical looses its green colour and becomes brown or yellow.