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DSMZ

DSMZ 585 . BCYE AGAR

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* Medium for Legionella oakridgensis DSM 21215

Humid

Growth conditions: 37 °C
Equipment needed: Autoclave Filter
Final pH: n.d.
Select a strain (optional):

ml
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 For solubilization of the lyophilized pellet use standard liquid media. Although Legionellae do not grow in those media they are suitable for resuspension and transfer onto agar plates.
4 Note: Alternatively use CYE-ACES AGAR

CYE-ACES AGAR #

1000
Compound Amount Unit
Solution A 490 ml
Solution B 490 ml
Solution C 10 ml
Solution D 10 ml
1 Add Cysteine-HCl, then ferric pyrophosphate solution to solution A. Adjust the complete medium to pH 6.9 ± 0.5 at 50°C with sterile 1.0 N KOH or HCl. The pH of the medium is critical. Finally, add solution B. Swirl medium in flask during dispensing to petri dishes or tubes in order to keep charcoal suspended.

Solution A #

490
Compound Amount Unit
Yeast extract
(Difco 0127) 		10 g
ACES
(N-2-acetamido-2-aminoethane-sulfonic acid) 		10 g
Activated charcoal
(Sigma C 5510) 		2 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.