Hydrothermal Vents: The Inhabitants, Their Way of Life and Their Adaptation to High Pressure

ABSTRACT We present a comparative analysis of two genome fragments isolated from a diverse and widely distributed group of uncultured euryarchaea from deep-sea hydrothermal vents. The optimal activity and thermostability of a DNA polymerase predicted in one fragment were close to that of the thermophilic archaeon Thermoplasma acidophilum, providing evidence for a thermophilic way of life of this group of uncultured archaea.

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Permanent ectoplasmic structures in deep-sea Cibicides/oides taxa – long-term observations at in situ pressure

10.5194/bg-2021-62 ◽

2021 ◽

Author(s):

Jutta Wollenburg ◽

Jelle Bijma ◽

Charlotte Cremer ◽

Ulf Bickmeyer ◽

Zora Mila Colomba Zittier

Keyword(s):

High Pressure ◽

Water Body ◽

Deep Sea ◽

Cell Structure ◽

Way Of Life ◽

Acetoxymethyl Ester ◽

Different Types ◽

Calcein Am

Abstract. Deep-sea Cibicidoides pachyderma (forma mundulus) and related Cibicidoides spp. were cultured at in situ pressure for 1-2 days, or 6 weeks to 3 months. During that period, fluorescence analyses following BCECF-AM (2’,7’-bis(2-carboxyethyl)-5-(and-6)-carboxyfluorescein acetoxymethyl ester) or Calcein AM (4,5-Bis((N,N-bis(carboxymethy)amino)methyl)fluorescein acetoxymethylester) labelling, revealed a persisting cytoplasmic sheet or envelope surrounding the Cibicidoides tests. Thus, the Cibicidoides shell can be considered rather as an internal than an external cell structure. A couple of days to a week after being transferred into high-pressure aquaria and adjusted to a pressure of 115 bar, the foraminifera changed from a mobile to a more or less sessile living mode. During this quasi sessile way of life, a series of comparably thick static ectoplasmic structures developed that were not resorbed or remodelled but, except for occasional further growth, remained unchanged throughout the experiments. Three different types of these ‘permanent structures’ were observed: A) Ectoplasmic ‘roots’ were common in adult C. pachyderma, C. lobatulus and C. wuellerstorfi specimens. In our experiments single ectoplasmic ‘roots’ grew to maximum 700 times the individuals shell diameter and were presumably used to anchor the specimen in an environment with strong currents. B) Ectoplasmic ‘trees’ describe rigid ectoplasmic structures directed into the aquarium’s water body and were used by the foraminifera to climb up and down these ectoplasmic structures. Ectoplasmic ‘trees’ were so far only observed in C. pachyderma and enabled the ‘tree’-forming foraminifera to elevate itself above ground. C) Ectoplasmic ‘twigs’ were used to guide and hold the more delicate pseudopodial network when distributed into prevailing currents, and were, in our experiments, also only developed in C. pachyderma specimens. Relocation of a specimen usually required to tear apart and leave behind the rigid ectoplasmic structures, eventually also the envelope surrounding the test. Apparently, these rigid structures could not be resorbed or reused.

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Chemosynthesis in the deep-sea: life without the sun

Biogeosciences Discussions ◽

10.5194/bgd-9-17037-2012 ◽

2012 ◽

Vol 9 (12) ◽

pp. 17037-17052 ◽

Cited By ~ 6

Author(s):

C. Smith

Keyword(s):

Organic Matter ◽

Surface Waters ◽

Deep Sea ◽

Hydrothermal Vents ◽

Cold Seep ◽

Cold Seeps ◽

The Sun ◽

Life Stages ◽

Way Of Life ◽

Use Of Resources

Abstract. Chemosynthetic communities in the deep-sea can be found at hydrothermal vents, cold seeps, whale falls and wood falls. While these communities have been suggested to exist in isolation from solar energy, much of the life associated with them relies either directly or indirectly on photosynthesis in the surface waters of the oceans. The sun indirectly provides oxygen, a byproduct of photosynthesis, which aerobic chemosynthetic microorganisms require to synthesize organic carbon from CO2. Planktonic life stages of many vent and cold seep invertebrates also directly feed on photosynthetically produced organic matter as they disperse to new vent and seep systems. While a large portion of the life at deep-sea chemosynthetic habitats can be linked to the sun and so could not survive without it, a small portion of anaerobically chemosynthetic microorganisms can persist in its absence. These small and exotic organisms have developed a way of life in the deep-sea which involves the use of resources originating in their entirety from terrestrial sources.

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

10.20944/preprints202104.0453.v1 ◽

2021 ◽

Author(s):

Gina Carole Oliver ◽

Anaïs Cario ◽

Karyn Lynne Rogers

Keyword(s):

High Pressure ◽

High Temperature ◽

Deep Sea ◽

Hydrothermal Vents ◽

Batch Cultivation ◽

Pressure Vessels ◽

Marine Microorganisms ◽

Thermophilic Microorganisms ◽

Variable Volume ◽

In Situ Conditions

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.

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Permanent ectoplasmic structures in deep-sea <i>Cibicides</i> and <i>Cibicidoides</i> taxa – long-term observations at in situ pressure

Biogeosciences ◽

10.5194/bg-18-3903-2021 ◽

2021 ◽

Vol 18 (12) ◽

pp. 3903-3915

Author(s):

Jutta E. Wollenburg ◽

Jelle Bijma ◽

Charlotte Cremer ◽

Ulf Bickmeyer ◽

Zora Mila Colomba Zittier

Keyword(s):

High Pressure ◽

Water Body ◽

Deep Sea ◽

Cell Structure ◽

Way Of Life ◽

Acetoxymethyl Ester ◽

Shell Diameter ◽

Different Types

Abstract. Deep-sea Cibicidoides pachyderma (forma mundulus) and related Cibicidoides spp. were cultured at in situ pressure for 1–2 d, or 6 weeks to 3 months. During that period, fluorescence analyses following BCECF-AM (2′,7′-bis(2-carboxyethyl)-5-(and-6)-carboxyfluorescein acetoxymethyl ester) or calcein (bis[N,N-bis(carboxymethyl)aminomethyl]-fluorescein) labelling revealed a persisting cytoplasmic sheet or envelope surrounding the Cibicidoides tests. Thus, the Cibicidoides shell can be considered as an internal rather than an external cell structure. A couple of days to a week after being transferred into high-pressure aquaria and adjusted to a pressure of 115 bar, the foraminifera changed from a mobile to a more or less sessile living mode. During this quasi-sessile way of life, a series of comparably thick static ectoplasmic structures developed that were not resorbed or remodelled but, except for occasional further growth, remained unchanged throughout the experiments. Three different types of these permanent structures were observed. (a) Ectoplasmic “roots” were common in adult C. pachyderma, C. lobatulus, and C. wuellerstorfi specimens. In our experiments single ectoplasmic roots grew to a maximum of 700 times the individuals' shell diameter and were presumably used to anchor the specimen in an environment with strong currents. (b) Ectoplasmic “trees” describe rigid ectoplasmic structures directed into the aquarium's water body and were used by the foraminifera to climb up and down these ectoplasmic structures. Ectoplasmic trees have so far only been observed in C. pachyderma and enabled the tree-forming foraminifera to elevate itself above ground. (c) Ectoplasmic “twigs” were used to guide and hold the more delicate pseudopodial network when distributed into prevailing currents and were, in our experiments, also only developed in C. pachyderma specimens. Relocation of a specimen usually required it to tear apart and leave behind the rigid ectoplasmic structures and eventually also the envelope surrounding the test. Apparently, these rigid structures could not be resorbed or reused.

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High-pressure freezing of cells in suspension for low-voltage scanning electron microscopy (LVSEM)

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100084624 ◽

1991 ◽

Vol 49 ◽

pp. 64-65

Author(s):

Marek Malecki ◽

James Pawley ◽

Hans Ris

Keyword(s):

Electron Microscopy ◽

High Pressure ◽

Low Voltage ◽

Culture Media ◽

Cell Structure ◽

Freeze Substitution ◽

Ice Crystal ◽

High Pressure Freezing ◽

Cell Culture Media ◽

Voltage Scanning

The ultrastructure of cells suspended in physiological fluids or cell culture media can only be studied if the living processes are stopped while the cells remain in suspension. Attachment of living cells to carrier surfaces to facilitate further processing for electron microscopy produces a rapid reorganization of cell structure eradicating most traces of the structures present when the cells were in suspension. The structure of cells in suspension can be immobilized by either chemical fixation or, much faster, by rapid freezing (cryo-immobilization). The fixation speed is particularly important in studies of cell surface reorganization over time. High pressure freezing provides conditions where specimens up to 500μm thick can be frozen in milliseconds without ice crystal damage. This volume is sufficient for cells to remain in suspension until frozen. However, special procedures are needed to assure that the unattached cells are not lost during subsequent processing for LVSEM or HVEM using freeze-substitution or freeze drying. We recently developed such a procedure.

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Prolonged Fixation Studies For Spaceflight

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100072101 ◽

1973 ◽

Vol 31 ◽

pp. 332-333

Author(s):

Robert Corbett ◽

Delbert E. Philpott ◽

Sam Black

Keyword(s):

High Pressure ◽

Living Systems ◽

Prolonged Storage ◽

Time Intervals ◽

Early Signs ◽

Large Numbers

Observation of subtle or early signs of change in spaceflight induced alterations on living systems require precise methods of sampling. In-flight analysis would be preferable but constraints of time, equipment, personnel and cost dictate the necessity for prolonged storage before retrieval. Because of this, various tissues have been stored in fixatives and combinations of fixatives and observed at various time intervals. High pressure and the effect of buffer alone have also been tried.Of the various tissues embedded, muscle, cartilage and liver, liver has been the most extensively studied because it contains large numbers of organelles common to all tissues (Fig. 1).

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Cryosectioning plant roots prepared by high-pressure freezing

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100127748 ◽

1987 ◽

Vol 45 ◽

pp. 662-663

Author(s):

R.E. Crang ◽

M. Mueller ◽

K. Zierold

Keyword(s):

High Pressure ◽

Cell Walls ◽

Plant Tissues ◽

Ice Crystal ◽

High Pressure Freezing ◽

Vitreous State ◽

Radial Depth ◽

Irregular Topography ◽

Considerable Depth ◽

Conventional Freezing

Obtaining frozen-hydrated sections of plant tissues for electron microscopy and microanalysis has been considered difficult, if not impossible, due primarily to the considerable depth of effective freezing in the tissues which would be required. The greatest depth of vitreous freezing is generally considered to be only 15-20 μm in animal specimens. Plant cells are often much larger in diameter and, if several cells are required to be intact, ice crystal damage can be expected to be so severe as to prevent successful cryoultramicrotomy. The very nature of cell walls, intercellular air spaces, irregular topography, and large vacuoles often make it impractical to use immersion, metal-mirror, or jet freezing techniques for botanical material.However, it has been proposed that high-pressure freezing (HPF) may offer an alternative to the more conventional freezing techniques, inasmuch as non-cryoprotected specimens may be frozen in a vitreous, or near-vitreous state, to a radial depth of at least 0.5 mm.

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High-pressure freezing and freeze substitution of plant tissue

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100124148 ◽

1992 ◽

Vol 50 (1) ◽

pp. 748-749

Author(s):

William P. Sharp ◽

Robert W. Roberson

Keyword(s):

High Pressure ◽

Cell Structure ◽

Leaf Tissue ◽

Freeze Substitution ◽

Crystal Formation ◽

Ice Crystal ◽

High Pressure Freezing ◽

Cell Components ◽

Cold Metal ◽

Brome Grass

The aim of ultrastructural investigation is to analyze cell architecture and relate a functional role(s) to cell components. It is known that aqueous chemical fixation requires seconds to minutes to penetrate and stabilize cell structure which may result in structural artifacts. The use of ultralow temperatures to fix and prepare specimens, however, leads to a much improved preservation of the cell’s living state. A critical limitation of conventional cryofixation methods (i.e., propane-jet freezing, cold-metal slamming, plunge-freezing) is that only a 10 to 40 μm thick surface layer of cells can be frozen without distorting ice crystal formation. This problem can be allayed by freezing samples under about 2100 bar of hydrostatic pressure which suppresses the formation of ice nuclei and their rate of growth. Thus, 0.6 mm thick samples with a total volume of 1 mm3 can be frozen without ice crystal damage. The purpose of this study is to describe the cellular details and identify potential artifacts in root tissue of barley (Hordeum vulgari L.) and leaf tissue of brome grass (Bromus mollis L.) fixed and prepared by high-pressure freezing (HPF) and freeze substitution (FS) techniques.

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Microstructural Study of Diamond Deformed Under High Pressure and Temperature

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100118072 ◽

1985 ◽

Vol 43 ◽

pp. 230-231

Author(s):

E. F. Koch

Keyword(s):

High Pressure ◽

High Temperature ◽

Lattice Structure ◽

Diamond Particle ◽

Polycrystalline Diamond ◽

Sintering Process ◽

Ion Milling ◽

Sintered Compact ◽

Transmission Electron ◽

High Pressure Sintering

Because of the extremely rigid lattice structure of diamond, generating new dislocations or moving existing dislocations in diamond by applying mechanical stress at ambient temperature is very difficult. Analysis of portions of diamonds deformed under bending stress at elevated temperature has shown that diamond deforms plastically under suitable conditions and that its primary slip systems are on the ﹛111﹜ planes. Plastic deformation in diamond is more commonly observed during the high temperature - high pressure sintering process used to make diamond compacts. The pressure and temperature conditions in the sintering presses are sufficiently high that many diamond grains in the sintered compact show deformed microtructures.In this report commercially available polycrystalline diamond discs for rock cutting applications were analyzed to study the deformation substructures in the diamond grains using transmission electron microscopy. An individual diamond particle can be plastically deformed in a high pressure apparatus at high temperature, but it is nearly impossible to prepare such a particle for TEM observation, since any medium in which the diamond is mounted wears away faster than the diamond during ion milling and the diamond is lost.

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