OLIVER, Gina Carole; CARIO, Anaïs; ROGERS, Karyn Lynne

doi:10.20944/preprints202104.0453.v1

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OLIVER, Gina Carole
Department of Earth and Environmental Sciences [Troy, NY]

CARIO, Anaïs
Department of Earth and Environmental Sciences [Troy, NY]

ROGERS, Karyn Lynne
Department of Earth and Environmental Sciences [Troy, NY]
Rensselaer Astrobiology Research and Education Center

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High temperatures (HT) and high hydrostatic pressures (HHP) are characteristic of deep-sea hydrothermal vents and other deep crustal settings. These environments host vast and diverse microbial populations, yet only a small fraction of those populations have been successfully cultured. This is due, in part, to the difficulty of sampling while maintaining these in situ conditions and also replicating those high-temperature and high-pressure conditions in the laboratory. In an effort to facilitate more HT-HHP cultivation, we present two HT-HHP batch culture incubation systems for cultivating deep-sea vent and subsurface (hyper)thermophilic microorganisms. One HT-HHP system can be used for batch cultivation up to 110 MPa and 121°C, and requires sample decompression during subsampling. The second HT-HHP system can be used to culture microorganisms up to 100 MPa and 160°C with variable-volume, pressure-retaining vessels that negate whole-sample decompression during subsampling. Here, we describe how to build cost effective heating systems for these two types of high-pressure vessels, as well as the protocols for HT-HHP microbial batch cultivation in both systems. Additionally, we demonstrate HHP transfer between the variable-volume vessels, which has utility in sampling and enrichment without decompression, laboratory isolation experiments, as well as HHP filtration.< Réduire

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High Temperature and High Hydrostatic Pressure Cultivation, Transfer, and Filtration Systems for Investigating Deep Marine Microorganisms

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