Leptospira alexanderi Brenner et al.

1. TRANSFER IMMEDIATELY upon thawing by aseptically withdrawing cell suspension from vial and inoculating a 10-12 mL tube of Medium #1470, inserting the pipette just below the surface of the semi-solid medium. Aliquots of 0.5 mL from this tube may be used to inoculate additional tubes if needed. An aerobic blood plate may also be streaked to test for purity.
2. Incubate test tubes with screw caps lightly tightened along with the blood plate at 30°C under aerobic conditions.
3. Initial growth may take up to 7-10 days. Observe growth as a thin, tight band forming just below the surface of the medium. The band thickens as incubation continues. 
4. The aerobic blood plate should show no signs of growth.

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