Acidithiobacillus ferrooxidans (Temple and Colmer) Kelly and Wood

1. Incubate test tube cultures under above conditions upon receipt.  It is helpful to incubate test tubes in a slanted position to increase gas exchange in broth.  Transfer culture to fresh media within one week of arrival.
2. Gently vortex the test-tube to dislodge the iron that has oxidized onto the glass.  Aseptically withdraw approximately 1.0 mL of the broth culture and transfer into 5 mL of fresh broth.
3. Incubate the broth in a static and slanted position.
4. Growth is evident within one to two weeks, when yellow-orange iron oxide deposits are observed.
5. Transfer the culture approximately every two to four weeks.
6. Transfer more tubes than needed as a backup source in the event of low viability due to repeated passage.  The culture will remain viable for a minimum of one month and for up to three months when stored at room temperature without shaking.

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