Culturing The Deep Subsurface

Natalia Obrzut, Caitlin Casar, and Magdalena Osburn
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Environmental fluid was used as inoculum in batch cultivation experiments under a variety of growth conditions [7]. Strains were isolated via serial transfers (Fig 5a) or dilution to extinction in cases where growth was not possible on solid media (Fig. 5b). Organisms Cultivated from DeMMO 1 um

Heterotrophic Growth Media Design.

Aerobic- Yeast peptone (YP) media was prepared with and without sulfate.

Anaerobic with Nitrate- YP media without sulfate was prepared anaerobically under a nitrogen headspace and supplemented with nitrate.

Anaerobic with Sulfate- YP media with sulfate was prepared anaerobically under a nitrogen headspace.

Autotrophic Growth Media Design.

Microaerobic with Ammonia- Base media without sulfate was supplemented with bicarbonate and ammonia. We targeted Archaea by suppressing bacterial growth with ampicillin. Cultures were prepared anaerobically under a nitrogen headspace and injected with filtered air.

Anaerobic with Ferrous Iron- Base media without sulfate was prepared anaerobically under a hydrogen headspace and supplemented with bicarbonate and magnetite or ferrihydrite.

Natalia Obrzut
Loyola University Chicago