scholarly journals Unexpected host dependency of Antarctic Nanohaloarchaeota

2019 ◽  
Vol 116 (29) ◽  
pp. 14661-14670 ◽  
Author(s):  
Joshua N. Hamm ◽  
Susanne Erdmann ◽  
Emiley A. Eloe-Fadrosh ◽  
Allegra Angeloni ◽  
Ling Zhong ◽  
...  
Keyword(s):  
Fluorescent In Situ Hybridization ◽  
Single Species ◽  
Hypersaline Environments ◽  
Free Living ◽  
Metagenome Analysis ◽  
Symbiotic Relationships ◽  
Cell Structures ◽  
Transmission Electron ◽  
Globally Distributed

In hypersaline environments, Nanohaloarchaeota (Diapherotrites, Parvarchaeota, Aenigmarchaeota, Nanoarchaeota, Nanohaloarchaeota [DPANN] superphylum) are thought to be free-living microorganisms. We report cultivation of 2 strains of Antarctic Nanohaloarchaeota and show that they require the haloarchaeon Halorubrum lacusprofundi for growth. By performing growth using enrichments and fluorescence-activated cell sorting, we demonstrated successful cultivation of Candidatus Nanohaloarchaeum antarcticus, purification of Ca. Nha. antarcticus away from other species, and growth and verification of Ca. Nha. antarcticus with Hrr. lacusprofundi; these findings are analogous to those required for fulfilling Koch’s postulates. We use fluorescent in situ hybridization and transmission electron microscopy to assess cell structures and interactions; metagenomics to characterize enrichment taxa, generate metagenome assembled genomes, and interrogate Antarctic communities; and proteomics to assess metabolic pathways and speculate about the roles of certain proteins. Metagenome analysis indicates the presence of a single species, which is endemic to Antarctic hypersaline systems that support the growth of haloarchaea. The presence of unusually large proteins predicted to function in attachment and invasion of hosts plus the absence of key biosynthetic pathways (e.g., lipids) in metagenome assembled genomes of globally distributed Nanohaloarchaeota indicate that all members of the lineage have evolved as symbionts. Our work expands the range of archaeal symbiotic lifestyles and provides a genetically tractable model system for advancing understanding of the factors controlling microbial symbiotic relationships.

scholarly journals Novel Parachlamydia acanthamoebae Quantification Method Based on Coculture with Amoebae

2008 ◽  
Vol 74 (20) ◽  
pp. 6397-6404 ◽  
Author(s):  
Junji Matsuo ◽  
Yasuhiro Hayashi ◽  
Shinji Nakamura ◽  
Marie Sato ◽  
Yoshihiko Mizutani ◽  
...  
Keyword(s):  
Fluorescent In Situ Hybridization ◽  
Vero Cells ◽  
Host Cells ◽  
Human Pathogens ◽  
Free Living ◽  
Infectious Dose ◽  
Quantification Method ◽  
Transmission Electron ◽  
Dilution Curves

ABSTRACT Parachlamydia acanthamoebae, belonging to the order Chlamydiales, is an obligately intracellular bacterium that infects free-living amoebae and is a potential human pathogen. However, no method exists to accurately quantify viable bacterial numbers. We present a novel quantification method for P. acanthamoebae based on coculture with amoebae. P. acanthamoebae was cultured either with Acanthamoeba spp. or with mammalian epithelial HEp-2 or Vero cells. The infection rate of P. acanthamoebae (amoeba-infectious dose [AID]) was determined by DAPI (4′,6-diamidino-2-phenylindole) staining and was confirmed by fluorescent in situ hybridization. AIDs were plotted as logistic sigmoid dilution curves, and P. acanthamoebae numbers, defined as amoeba-infectious units (AIU), were calculated. During culture, amoeba numbers and viabilities did not change, and amoebae did not change from trophozoites to cysts. Eight amoeba strains showed similar levels of P. acanthamoebae growth, and bacterial numbers reached ca. 1,000-fold (109 AIU preculture) after 4 days. In contrast, no increase was observed for P. acanthamoebae in either mammalian cell line. However, aberrant structures in epithelial cells, implying possible persistent infection, were seen by transmission electron microscopy. Thus, our method could monitor numbers of P. acanthamoebae bacteria in host cells and may be useful for understanding chlamydiae present in the natural environment as human pathogens.

scholarly journals Microsporidian Infection in a Free-Living Marine Nematode

2012 ◽  
Vol 11 (12) ◽  
pp. 1544-1551 ◽  
Author(s):  
A. M. Ardila-Garcia ◽  
N. M. Fast
Keyword(s):  
Marine Nematode ◽  
Free Living ◽  
Content Type ◽  
Infection Pattern ◽  
Transmission Electron ◽  
Muscle Tissues ◽  
Natural Hosts ◽  
In The Wild ◽  
Microsporidian Infection

ABSTRACT Microsporidia are unicellular fungi that are obligate endoparasites. Although nematodes are one of the most abundant and diverse animal groups, the only confirmed report of microsporidian infection was that of the “nematode killer” ( Nematocida parisii ). N. parisii was isolated from a wild Caenorhabditis sp. and causes an acute and lethal intestinal infection in a lab strain of Caenorhabditis elegans . We set out to characterize a microsporidian infection in a wild nematode to determine whether the infection pattern of N. parisii in the lab is typical of microsporidian infections in nematodes. We describe a novel microsporidian species named Sporanauta perivermis (marine spore of roundworms) and characterize its infection in its natural host, the free-living marine nematode Odontophora rectangula. S. perivermis is not closely related to N. parisii and differs strikingly in all aspects of infection. Examination by transmission electron microscopy (TEM) revealed that the infection was localized in the hypodermal and muscle tissues only and did not involve the intestines. Fluorescent in situ hybridization (FISH) confirmed infection in the muscle and hypodermis, and surprisingly, it also revealed that the parasite infects O. rectangula eggs, suggesting a vertical mode of transmission. Our observations highlight the importance of studying parasites in their natural hosts and indicate that not all nematode-infecting microsporidia are “nematode killers”; instead, microsporidiosis can be more versatile and chronic in the wild.

Transitions in symbiosis: evidence for environmental acquisition & social transmission within a clade of heritable symbionts

2020 ◽  
Author(s):  
Georgia C Drew ◽  
Giles E Budge ◽  
Crystal L Frost ◽  
Peter Neumann ◽  
Stefanos Siozios ◽  
...  
Keyword(s):  
Honey Bee ◽  
Honey Bees ◽  
Fluorescent In Situ Hybridization ◽  
Horizontal Transmission ◽  
Bacterial Symbionts ◽  
Free Living ◽  
Social Contacts ◽  
Obligate Mutualism ◽  
Environmental Microbes

AbstractA dynamic continuum exists from free-living environmental microbes to strict host associated symbionts that are vertically inherited. However, knowledge of the forces that drive transitions in the modes by which symbioses form is lacking. Arsenophonus is a diverse clade of bacterial symbionts, comprising reproductive parasites to coevolving obligate mutualists, in which the predominant mode of transmission is vertical. We describe a symbiosis between a member of the genus Arsenophonus and the Western honey bee. We then present multiple lines of evidence that this symbiont deviates from a heritable model of transmission. Field sampling uncovered marked spatial and seasonal dynamics in symbiont prevalence, and rapid infection loss events were observed in field colonies and individuals in the laboratory. Fluorescent in-situ hybridization showed Arsenophonus localised in the gut, and detection of the bacterium was rare in screens of early honey bee life stages. We directly show horizontal transmission of Arsenophonus between bees under varying social conditions. We conclude that honey bees acquire Arsenophonus through a combination of environmental exposure and social contacts. Together these findings uncover a key link in the Arsenophonus clades trajectory from free-living ancestral life to obligate mutualism, and provide a foundation for studying transitions in symbiotic lifestyle.

scholarly journals A Single Species, Micromonas pusilla (Prasinophyceae), Dominates the Eukaryotic Picoplankton in the Western English Channel

2004 ◽  
Vol 70 (7) ◽  
pp. 4064-4072 ◽  
Author(s):  
Fabrice Not ◽  
Mikel Latasa ◽  
Dominique Marie ◽  
Thierry Cariou ◽  
Daniel Vaulot ◽  
...  
Keyword(s):  
Aquatic Ecosystems ◽  
Fluorescent In Situ Hybridization ◽  
Algal Species ◽  
Sampling Period ◽  
Single Species ◽  
English Channel ◽  
Green Algal ◽  
Micromonas Pusilla ◽  
Green Algal Species

ABSTRACT The class Prasinophyceae (Chlorophyta) contains several photosynthetic picoeukaryotic species described from cultured isolates. The ecology of these organisms and their contributions to the picoeukaryotic community in aquatic ecosystems have received little consideration. We have designed and tested eight new 18S ribosomal DNA oligonucleotide probes specific for different Prasinophyceae clades, genera, and species. Using fluorescent in situ hybridization associated with tyramide signal amplification, these probes, along with more general probes, have been applied to samples from a marine coastal site off Roscoff (France) collected every 2 weeks between July 2000 and September 2001. The abundance of eukaryotic picoplankton remained high (>103 cells ml−1) during the sampling period, with maxima in summer (up to 2 × 104 cells ml−1), and a single green algal species, Micromonas pusilla (Prasinophyceae), dominated the community all year round. Members of the order Prasinococcales and the species Bathycoccus prasinos (Mamiellales) displayed sporadic occurrences, while the abundances of all other Prasinophyceae groups targeted remained negligible.

The Use of Advanced Analytical Techniques for Characterization of the Chemical, Physical, and Crystallographic Nature of a Surface

1973 ◽  
Vol 31 ◽  
pp. 132-133 ◽  
Author(s):  
R. E. Herfert
Keyword(s):  
Surface Characterization ◽  
Analytical Techniques ◽  
X Ray Diffraction ◽  
Depth Of Penetration ◽  
X Ray ◽  
The Past ◽  
Transmission Electron ◽  
Scanning Electron

Studies of the nature of a surface, either metallic or nonmetallic, in the past, have been limited to the instrumentation available for these measurements. In the past, optical microscopy, replica transmission electron microscopy, electron or X-ray diffraction and optical or X-ray spectroscopy have provided the means of surface characterization. Actually, some of these techniques are not purely surface; the depth of penetration may be a few thousands of an inch. Within the last five years, instrumentation has been made available which now makes it practical for use to study the outer few 100A of layers and characterize it completely from a chemical, physical, and crystallographic standpoint. The scanning electron microscope (SEM) provides a means of viewing the surface of a material in situ to magnifications as high as 250,000X.

In situ REM imaging of surface processes on ceramic bulk crystals from 300 to 1670 K in a conventional TEM

1991 ◽  
Vol 49 ◽  
pp. 660-661
Author(s):  
Z. L. Wang ◽  
J. Bentley
Keyword(s):  
High Temperatures ◽  
Image Resolution ◽  
Atomic Processes ◽  
Crystal Surfaces ◽  
Beam Radiation ◽  
Surface Areas ◽  
Transmission Electron ◽  
Reflection Electron Microscopy ◽  
Gas Reactions

Studying the behavior of surfaces at high temperatures is of great importance for understanding the properties of ceramics and associated surface-gas reactions. Atomic processes occurring on bulk crystal surfaces at high temperatures can be recorded by reflection electron microscopy (REM) in a conventional transmission electron microscope (TEM) with relatively high resolution, because REM is especially sensitive to atomic-height steps.Improved REM image resolution with a FEG: Cleaved surfaces of a-alumina (012) exhibit atomic flatness with steps of height about 5 Å, determined by reference to a screw (or near screw) dislocation with a presumed Burgers vector of b = (1/3)<012> (see Fig. 1). Steps of heights less than about 0.8 Å can be clearly resolved only with a field emission gun (FEG) (Fig. 2). The small steps are formed by the surface oscillating between the closely packed O and Al stacking layers. The bands of dark contrast (Fig. 2b) are the result of beam radiation damage to surface areas initially terminated with O ions.

Role of boundaries in the crystallization of amorphous films

1988 ◽  
Vol 46 ◽  
pp. 626-627
Author(s):  
D. A. Smith
Keyword(s):  
Low Temperature ◽  
Amorphous State ◽  
Nucleation And Growth ◽  
Amorphous Films ◽  
Island Nucleation ◽  
Surface Steps ◽  
Transmission Electron ◽  
Component Systems ◽  
Temperature Deposition

The nucleation and growth processes which lead to the formation of a thin film are particularly amenable to investigation by transmission electron microscopy either in situ or subsequent to deposition. In situ studies have enabled the observation of island nucleation and growth, together with addition of atoms to surface steps. This paper is concerned with post-deposition crystallization of amorphous alloys. It will be argued that the processes occurring during low temperature deposition of one component systems are related but the evidence is mainly indirect. Amorphous films result when the deposition conditions such as low temperature or the presence of impurities (intentional or unintentional) preclude the atomic mobility necessary for crystallization. Representative examples of this behavior are CVD silicon grown below about 670°C, metalloids, such as antimony deposited at room temperature, binary alloys or compounds such as Cu-Ag or Cr O2, respectively. Elemental metals are not stable in the amorphous state.

In situ observations of electromigration induced void behavior

1995 ◽  
Vol 53 ◽  
pp. 244-245
Author(s):  
T. Marieb ◽  
J. C. Bravman ◽  
P. Flinn ◽  
D. Gardner ◽  
M. Madden
Keyword(s):  
Electron Microscopy ◽  
Void Nucleation ◽  
Plan View ◽  
Transmission Electron ◽  
Nucleation Sites ◽  
Microelectronics Industry ◽  
In Situ Observations ◽  
Backscatter Electron Detector ◽  
Voltage Scanning

Electromigration and stress voiding have been active areas of research in the microelectronics industry for many years. While accelerated testing of these phenomena has been performed for the last 25 years[1-2], only recently has the introduction of high voltage scanning electron microscopy (HVSEM) made possible in situ testing of realistic, passivated, full thickness samples at high resolution.With a combination of in situ HVSEM and post-testing transmission electron microscopy (TEM) , electromigration void nucleation sites in both normal polycrystalline and near-bamboo pure Al were investigated. The effect of the microstructure of the lines on the void motion was also studied.The HVSEM used was a slightly modified JEOL 1200 EX II scanning TEM with a backscatter electron detector placed above the sample[3]. To observe electromigration in situ the sample was heated and the line had current supplied to it to accelerate the voiding process. After testing lines were prepared for TEM by employing the plan-view wedge technique [6].

A TEM study of the microstructure of avian eggshells

1992 ◽  
Vol 50 (2) ◽  
pp. 1102-1103
Author(s):  
S. Q. Xiao ◽  
S. Baden ◽  
A. H. Heuer
Keyword(s):  
Electron Microscope ◽  
Calcium Carbonate ◽  
Nucleation And Growth ◽  
Growth Mechanisms ◽  
Shell Surface ◽  
Thin Sections ◽  
Polycrystalline Metals ◽  
Transmission Electron ◽  
Nucleation And Growth Mechanisms

The avian eggshell is one of the most rapidly mineralizing biological systems known. In situ, 5g of calcium carbonate are crystallized in less than 20 hrs to fabricate the shell. Although there have been much work about the formation of eggshells, controversy about the nucleation and growth mechanisms of the calcite crystals, and their texture in the eggshell, still remain unclear. In this report the microstructure and microchemistry of avian eggshells have been analyzed using transmission electron microscope (TEM) and energy dispersive spectroscopy (EDS).Fresh white and dry brown eggshells were broken and fixed in Karnosky's fixative (kaltitanden) for 2 hrs, then rinsed in distilled H2O. Small speckles of the eggshells were embedded in Spurr medium and thin sections were made ultramicrotome.The crystalline part of eggshells are composed of many small plate-like calcite grains, whose plate normals are approximately parallel to the shell surface. The sizes of the grains are about 0.3×0.3×1 μm3 (Fig.l). These grains are not as closely packed as man-made polycrystalline metals and ceramics, and small gaps between adjacent grains are visible indicating the absence of conventional grain boundaries.

Dynamic studies of silicide-mediated crystallization of amorphous silicon

1992 ◽  
Vol 50 (2) ◽  
pp. 1352-1353
Author(s):  
C. Hayzelden ◽  
J. L. Batstone
Keyword(s):  
Ion Implantation ◽  
Solid Phase ◽  
Metallic Phase ◽  
Single Crystal Substrate ◽  
Free Electrons ◽  
Melt Temperature ◽  
Transmission Electron ◽  
Crystal Substrate ◽  
High Resolution Tem

Epitaxial reordering of amorphous Si(a-Si) on an underlying single-crystal substrate occurs well below the melt temperature by the process of solid phase epitaxial growth (SPEG). Growth of crystalline Si(c-Si) is known to be enhanced by the presence of small amounts of a metallic phase, presumably due to an interaction of the free electrons of the metal with the covalent Si bonds near the growing interface. Ion implantation of Ni was shown to lower the crystallization temperature of an a-Si thin film by approximately 200°C. Using in situ transmission electron microscopy (TEM), precipitates of NiSi2 formed within the a-Si film during annealing, were observed to migrate, leaving a trail of epitaxial c-Si. High resolution TEM revealed an epitaxial NiSi2/Si(l11) interface which was Type A. We discuss here the enhanced nucleation of c-Si and subsequent silicide-mediated SPEG of Ni-implanted a-Si.Thin films of a-Si, 950 Å thick, were deposited onto Si(100) wafers capped with 1000Å of a-SiO2. Ion implantation produced sharply peaked Ni concentrations of 4×l020 and 2×l021 ions cm−3, in the center of the films.

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