Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

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

Version 1 : Received: 15 April 2021 / Approved: 17 April 2021 / Online: 17 April 2021 (08:46:55 CEST)

How to cite: Oliver, G.C.; Cario, A.; Rogers, K.L. High Temperature and High Hydrostatic Pressure Cultivation, Transfer, and Filtration Systems for Investigating Deep Marine Microorganisms. Preprints 2021, 2021040453 (doi: 10.20944/preprints202104.0453.v1). Oliver, G.C.; Cario, A.; Rogers, K.L. High Temperature and High Hydrostatic Pressure Cultivation, Transfer, and Filtration Systems for Investigating Deep Marine Microorganisms. Preprints 2021, 2021040453 (doi: 10.20944/preprints202104.0453.v1).

Abstract

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.

Subject Areas

high-pressure microbiology; deep biosphere; cultivation devices; decompression

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