Microm In Width Research Articles

Morphological and ultrastructural redescription of Chloromyxum leydigi Mingazzini, 1890 (Myxozoa: Myxosporea), type species of the genus, infecting the gall bladder of the marine cartilaginous fish Torpedo marmorata Risso (Chondrichthyes: Torpedinidae), from the Portuguese Atlantic coast

Chloromyxum leydigi Mingazzini, 1890, the type species of Chloromyxum Mingazzini, 1890, is redescribed based on material found in the gall bladder of the cartilaginous fish Torpedo marmorata Risso collected from the Portuguese Atlantic coast and its sporogonic development is described. Plasmodia and mature spores were floating free in the bile. Plasmodia are polysporic and highly polymorphic in shape and organization. Mature spores are spherical to subspherical with a pointed anterior end, measuring 12.3 +/- 0.5 microm in length and 9.0 +/- 0.5 microm in width. The spore wall is composed of two asymmetric shell valves, each bearing 4-5 elevated surface ridges. A bundle of 40-50 tapering caudal filaments extends from the basal portion of the shell valves. Four pyriform equal-sized polar capsules, measuring about 5.3 x 3.2 microm, are observed at the same level in the anterior pole of the spores, each containing a polar filament coiled in 8-9 (rarely 10) turns. Spore morphology, tissue tropism, host species and sequences of the SSU rRNA gene supported species identification. Since its discovery, this species has been dubiously reported from several cartilaginous hosts, namely due to the poor description of its features.

Aggregata bathytherma sp. nov. (Apicomplexa: Aggregatidae), a new coccidian parasite associated with a deep-sea hydrothermal vent octopus

Aggregata bathytherma sp. nov. is described from the digestive tract of Vulcanoctopus hydrothermalis, a deep-sea octopus recently discovered associated with hydrothermal vents in the northeast Pacific Ocean. Oocysts typically are spherical in shape, sometimes irregular, 163 to 356 microm in length, and 219 to 313 microm in width. Each oocyst contains from 50 to over 200 sporocysts. Sporocysts measure 27 to 32 microm in longest diameter. The cyst wall is smooth and 1 microm thick. Each sporocyst typically contains 14 to 17 sporozoites, 49 microm in length. Histological lesions associated with the presence of A. bathytherma include rupture of the basal membrane and detachment of the epithelial cells. In heavily infected areas, most of the tissue of the host digestive tract is replaced by parasites. A. bathytherma is the first Aggregata species described from a host that lives in association with hydrothermal vents, and the third species of Aggregata from eastern North Pacific waters.

Transfer-Free Batch Fabrication of Large-Area Suspended Graphene Membranes

We demonstrate a process for batch production of large-area (100-3000 microm(2)) patterned free-standing graphene membranes on Cu scaffolds using chemical vapor deposition (CVD)-grown graphene. This technique avoids the use of silicon and transfers of graphene. As one application of this technique, we fabricate transmission electron microscopy (TEM) sample supports. TEM characterization of the graphene membranes reveals relatively clean, highly TEM-transparent, single-layer graphene regions ( approximately 50% by area) and, despite the polycrystalline nature of CVD graphene, membrane yields as high as 75-100%. This high yield verifies that the intrinsic strength and integrity of CVD-grown graphene films is sufficient for sub-100 microm width membrane applications. Elemental analysis (electron energy loss spectroscopy (EELS) and X-ray energy-dispersive spectroscopy (EDS)) of the graphene membranes reveals some nanoscaled contamination left over from the etching process, and we suggest several ways to reduce this contamination and improve the quality of the graphene for electronic device applications. This large-scale production of suspended graphene membranes facilitates access to the two-dimensional physics of graphene that are suppressed by substrate interactions and enables the widespread use of graphene-based sample supports for electron and optical microscopy.

Microstructured KY(WO_4)_2:Gd^3+, Lu^3+, Yb^3+ channel waveguide laser

Epitaxially grown, 2.4-microm-thin layers of KY(WO(4))(2):Gd(3+), Lu(3+), Yb(3+), which exhibit a high refractive index contrast with respect to the undoped KY(WO(4))(2) substrate, have been microstructured by Ar beam milling, providing 1.4-microm-deep ridge channel waveguides of 2 to 7 microm width, and overgrown by an undoped KY(WO(4))(2) layer. Channel waveguide laser operation was achieved with a launched pump power threshold of only 5 mW, a slope efficiency of 62% versus launched pump power, and 76 mW output power.

Copper-Ion-Assisted Self-Assembly of Silicate Clays in Rod- and Disklike Morphologies

Self-assembled rodlike (0.3-2.5 microm in diameter and 5.3-31 microm in length) and disklike microstructures (1.8-10.6 microm in width and 0.1-1.0 microm in thickness) are uniquely present in amorphous clay aggregates. Clay units were prepared by intercalation of Na(+)-montmorillonite (Na(+)-MMT) with copper ions (Cu(2+)) and poly(oxypropylene)-amine salt (POP) in simultaneous or stepwise ionic exchange reactions. Differences in process control during incorporation of Cu(2+) and hydrophobic POP greatly affected the layer structure of the clay units (d spacing of 12-53 A) and consequently their amphiphilic dispersion properties. By controlling the dispersion in water and drying at 80 degrees C, highly ordered self-assembly structures were obtained, presumably as a result of self-piling of clay units in competing vertical and horizontal directions. In general, association with Cu(2+) yielded units with a disklike microstructure, in contrast to the rod-like structure obtained for POP-intercalated clay. The self-assembled structures were characterized using X-ray diffraction, UV adsorption, thermal gravimetric analysis, zeta potential, scanning electron microscopy, and energy-dispersive X-ray spectroscopy techniques. Control of the clay self-piling process provides a new synthetic route for the fabrication of bottom-up microstructures that are potentially useful for templates, sensors, and electronic devices.

Clinical utility of antioxidant gene expression levels in circulating cancer cell clusters for the detection of prostate cancer in patients with prostate‐specific antigen levels of 4–10 ng/mL and disease prognostication after radical prostatectomy

To test antioxidant genes (AOX) expression in circulating cancer cell clusters (CCC). A novel method using molecular, polymerase chain reaction (PCR)-based detection of CCC was applied for predicting prostate cancer and to assess the effect of radical prostatectomy (RP) on reducing CCC and for prognostication of relapse-free survival (RFS), as serum total prostate-specific antigen (tPSA) has limited specificity at 4-10 ng/mL. In all, 240 patients were enrolled in the study, 129 for tumour diagnosis and 111 after RP for disease prognostication. Filtration assay in previously fractionated mononuclear cells (MNC) was used to enrich the CCC and large cells, which were retained in a mesh of 20 microm width. To establish the malignant nature of these cells they were analysed for genomic imbalances detected via PCR-assays of loss of heterozygosity in tumour suppressor loci and of DNA amplification in protooncogen loci. As a screening test in daily practice, real-time reverse transcription (RT)-PCR of AOX was introduced to overcome the laborious and expensive DNA tests. The AOX chosen were glutathione peroxidase (GPX1), Mn-dependent superoxide dismutase 2 (SOD2) and thioredoxine reductase (TXNRD1); selected from 67 marker candidate genes according to sensitivity and specificity data. AOX overexpression in CCC serves as a general marker for solid tumours needing, however, organ markers to relate to the organ of origin. Androgen receptor (AR), PSA and prostate-specific membrane antigen mRNAs served as organ markers for the prostate. Signals were detected in patients' MNC and to a minor level in CCC, rendering to CCC a substantial loss in epithelial features equivalent to a lower grade of epithelial differentiation. Organ markers in the MNC fraction were positive in <85% of AOX testing. The AOX test was tumour predicting (P < 0.001) with a sensitivity of 86%, specificity 82%, positive predictive value 69%, negative predictive value 92%, accuracy 83% and odds ratio (OR) of 28. SOD2 and TXNRD1 expression correlated to tumour size and Gleason score. Objective assessment for the evaluation of the molecular cell markers was achieved by receiver operating characteristic (ROC) curves. The areas under the ROC curve values of the AOXs were 0.7-0.9. RP was followed by a complete clearance of AOX-expressing cells. After RP, a subgroup of patients had residual CCC over-expressing only SOD2 and GPX1 indicating incomplete clearance by RP. Sustained overexpression of SOD2 and GPX1 accounted as risk factors for distant tumour recurrence (P = 0.003) mainly for bone metastases (97% M1b) as evaluated by Kaplan-Meier curves. In univariate analysis the tumour size had a limited effect on the probability of RFS (P = 0.05). In multivariate analysis tumour size, nodal status and Gleason score had no effect. This can partially be attributed to the higher risk level of pathological variables in the AOX over-expressing group but also to ineffective endocrine therapy resulting in marked overexpression of ARs and GPX1, the lead prognosticator gene. The AOX expression level allowed the identification of patients with high progression risk, who have more favourable pathological variables. The AOX testing of CCC is a novel method with excellent prognostic and predictive properties enabling the monitoring of therapies, e.g. effects of RP and endocrine therapy. We speculate that the continuing elevated expression of AOX with special emphasis on GPX1 acts as survival and defence mechanism in CCC required in an atypical environment prone to escape from immune surveillance.

Thin Films of Conductive ZnO Patterned by Micromolding Resulting in Nearly Isolated Features

Patterned and continuous thin films of conductive Al-doped zinc oxide (ZnO:Al) were prepared on different substrates from a polymeric precursor solution. Their electric conductivity and light transmittance (for visible and UV light) was measured at room temperature. By means of a simple device, conductive ZnO:Al films with high fidelity patterns with features of 2-20 microm width could be obtained by simply micromolding the liquid precursor film prior to heat treatment. The individual features were interconnected by a very thin residual ZnO layer.

Direct patterning of silicon oxide on Si-substrate induced by femtosecond laser

In this study we report for the first time a method for direct patterning of silicon oxide on a silicon substrate by irradiation with a femtosecond laser of Mega Hertz pulse frequency under ambient condition. Embossed lines of silicon oxide with around 3 approximately 4 microm width and less than 100 nm height were formed by controlling the parameters such as laser pulse power and frequency rate. A Scanning Electron Microscope (SEM), an optical microscopy and a Micro-Raman and Energy Dispersive X-ray (EDX) spectroscopy were used to analyze the silicon oxide layer.

Comparative Study on Histological Structures of the Vitelline Membrane of Hen and Duck Egg Observed by Cryo-Scanning Electron Microscopy

The histological structures of the vitelline membranes (VM) of hen and duck eggs were observed by cryo-scanning electron microscopy (cryo-SEM), and the chemical characteristics were also compared. The outer layer surface (OLS) of duck egg VM showed networks constructed by fibrils and sheets (0.1-5.2 microm in width), and that of hen egg presented networks formed only by sheets (2-6 microm in width). Thicker fibrils (0.5-1.5 microm in width) with different arrangement were observed on the inner layer surface (ILS) of duck egg VM as compared to those (0.3-0.7 microm in width) of hen egg VM. Upon separation, the outer surface of the outer layer (OSOL) and the inner surface of the inner layer (ISIL) of hen and duck egg VMs were quite similar to fresh VM except that the OSOL of duck egg VM showed networks constructed only by sheets. Thin fibrils interlaced above a bumpy or flat structure were observed at the exposed surface of the outer layer (ESOL) of hen and duck egg VMs. The exposed surfaces of inner layers (ESIL) of hen and duck egg VMs showed similar structures of fibrils, which joined, branched, and ran in straight lines for long distances up to 30 microm; however, the widths of the fibrils shown in ESOL and ESIL of duck egg VM were 0.1 and 0.7-1.4 microm, respectively, and were greater than those (<0.1 and 0.5-0.8 microm) of hen egg VM. The continuous membranes of both hen and duck egg VMs were still attached to the outer layers when separated. The content of protein, the major component of VM, was higher in duck egg VM (88.6%) than in hen egg VM (81.6%). Four and six major SDS-soluble protein patterns with distinct localization were observed in hen and duck egg VMs, respectively. Overall, the different histological structures of hen and duck egg VMs were suggested to be majorly attributable to the diverse protein components.

Low-cost rapid prototyping of flexible microfluidic devices using a desktop digital craft cutter

Low-cost and straight forward rapid prototyping of flexible microfluidic devices using a desktop digital craft cutter is presented. This rapid prototyping method can consistently achieve microchannels as thin as 200 microm in width and can be used to fabricate three-dimensional (3D) microfluidic devices using only double-sided pressure sensitive adhesive (PSA) tape and laser printer transparency film. Various functional microfluidic devices are demonstrated with this rapid prototyping method. The complete fabrication process from device design concept to working device can be completed in minutes without the need of expensive equipment.

Sarcocystis calchasi sp. nov. of the domestic pigeon (Columba livia f. domestica) and the Northern goshawk (Accipiter gentilis): light and electron microscopical characteristics

A novel highly pathogenic Sarcocystis species has been shown to cycle between the Northern goshawk (Accipiter gentilis) as definitive host and the domestic pigeon (Columba livia f. domestica) as intermediate host. However, genetically based characteristics are only available from very few bird-infecting Sarcocystis species. We therefore further characterised morphological properties of this protozoan in both hosts. Using light and electron microscopy, oocysts and sporocysts as well as schizonts and sarcocysts were characterised and compared with available morphological features of previously reported Sarcocystis species of Northern goshawks, Columbidae and genetically closely related species of other avian hosts. Sporocysts shed from day 6 on after experimental infection by the Northern goshawk were of ovoid appearance (11.9 x 7.9 microm). Ultrastructurally, schizonts of all developmental stages were found in the liver, spleen and next to or in endothelial cells of various organs of domestic pigeons 7 to 12 days after experimental infection. The cyst wall surface of slender sarcocysts (1 to 2 mm in length and 20 to 50 microm in width) was smooth and lacked protrusions. Cystozoites were lancet-shaped and measured 7.5 x 1.5 microm in Giemsa stain smears. The morphological findings, when combined with data of experimental infection and genetic studies, convergently indicate that the recently discovered Sarcocystis species represents a new species. We therefore propose to name this parasite Sarcocystis calchasi species nova.

Application of a simple asynchronous mechanical light chopper to multielectron coincidence spectroscopy

A simple asynchronous mechanical light chopper, based on modification of a turbo-molecular pump, has been developed to extend the interval between light pulses in single bunch operation at the Photon Factory storage ring. A pulse repetition rate of 80 kHz was achieved using a cylinder rotating at 48000 rpm, with 100 slits of 80 microm width. This allows absolute timing of particles up to 12.48 micros instead of the single-bunch period of 624 ns. We have applied the chopper together with a light pulse monitor to measure multielectron coincidence spectra using a magnetic bottle time-of-flight electron spectrometer. With such a system, the electron energies are determined without any ambiguity, the folding of coincidence spectra disappears and the effect of false coincidences is drastically reduced.

Transparent Conductive Coatings by Printing Coffee Ring Arrays Obtained at Room Temperature

We report here a concept for utilization of the "coffee ring effect" and inkjet printing to obtain transparent conductive patterns, which can replace the widely used transparent conductive oxides, such as ITO. The transparent conductive coating is achieved by forming a 2-D array of interconnected metallic rings. The rim of the individual rings is less than 10 microm in width and less than 300 nm in height, surrounding a "hole" with a diameter of about 150 microm; therefore the whole array of the interconnected rings is almost invisible to the naked eye. The rims of the rings are composed of self-assembled, closely packed silver nanoparticles, which make the individual rings and the resulting array electrically conductive. The resulting arrays of rings have a transparency of 95%; resistivity of 0.5 cm(2) was 4 +/- 0.5 Omega/, which is better than conventional ITO transparent thin films. The silver rings and arrays are fabricated by a very simple, low cost process, based on inkjet printing of a dispersion of 0.5 wt % silver nanoparticles (approximately 20 nm diameter) on plastic substrates. The performance of this transparent conductive coating was demonstrated by using it as an electrode for a plastic electroluminescent device, demonstrating the applicability of this concept in plastics electronics. It is expected that such transparent conductive coatings can be used in a wide range of applications such as displays (LCD, plasma, touch screens, e-paper), lighting devices (electroluminescence, OLED), and solar cells.

Insights into Advanced Retinopathy of Prematurity Using Handheld Spectral Domain Optical Coherence Tomography Imaging

To elucidate the subclinical anatomy of retinopathy of prematurity (ROP) using spectral domain optical coherence tomography (SD OCT). Prospective, observational case series. Three low-birth-weight, severely premature infants. Clinical examination was performed using a portable slit lamp and indirect ophthalmoscope. Imaging was performed by using a handheld SD OCT device and Retcam (Clarity Medical Systems, Pleasanton, CA) or video-indirect recording. Spectral domain optical coherence tomography imaging was conducted without sedation at the bedside in the neonatal intensive care unit on 1 patient. The other 2 patients had an examination under anesthesia with SD OCT imaging in the operating room. In vivo determination of vitreoretinal morphology, anatomy, and pathology by clinical examination, imaging, and SD OCT. Linear and volumetric imaging was achieved with the handheld system in infant eyes despite tunica vasculosa lentis and vitreous bands. Imaging was not possible in eyes with notable vitreous hemorrhage. Analysis of SD OCT images revealed preretinal structures (ranging from 409 to 2700 microm in width and 212 to 440 microm in height), retinoschisis, and retinal detachment in the posterior pole of patients with advanced ROP. Both the retinoschisis and the preretinal structures were not identified on conventional examination or imaging by expert pediatric ophthalmologists. The preretinal structures varied in location and size, and may represent preretinal fibrovascular proliferation. Some were found in close proximity to blood vessels, whereas others were near the optic nerve. Handheld SD OCT imaging can be performed on the sedated or nonsedated neonate and provides valuable subclinical anatomic information. This novel imaging modality can reveal the location and extent of posterior ROP pathology not evident on standard examination. This could affect future clinical decision-making if studies validate a management strategy based on findings from this imaging technique.

Exophiala xenobiotica infection in cultured striped jack, Pseudocaranx dentex (Bloch &amp; Schneider), in Japan

This report describes Exophiala infection in cultured striped jack, Pseudocaranx dentex, in Japan in 2005. One hundred out of 35,000 fish died per day and mortalities continued for 1 month. Diseased fish showed swelling of the abdomen and kidney distension. Numerous septate hyphae, pale brown in colour, were seen in kidney in squash preparations. Histology revealed abundant fungal hyphae and conidia in gill, heart and kidney. Fungal hyphae were accompanied by cell necrosis and influx of inflammatory, mainly mononuclear cells. The fungus isolated from the diseased fish had septate hyphae, pale brown in colour and 1.8-3.0 microm in diameter. Conidiogenous cells were conspicuous annellides, short or cylindrical or fusiform in shape. Conidia were one-celled, ellipsoidal with smooth walls, accumulated in balls at the apices of annellides that tended to slide down, 1.5-2.0 microm in width and 3.0-5.0 microm in length. The fungus was classified into the genus Exophiala based on its morphology and as Exophiala xenobiotica based on the sequences of the ITS 1-5.8S-ITS 2 regions of rDNA. This is the first record of this fungus in a marine fish.

Micropatterning of single myotubes on a thermoresponsive culture surface using elastic stencil membranes for single-cell analysis

We have developed a micropatterning procedure for single myotubes and demonstrated recovery of patterned myotubes without the use of methods that might cause damage to the cells. Since skeletal muscle is a highly ordered tissue mainly composed of myotubes, analysis of single myotubes is one of the promising approaches for studying the various diseases related to skeletal muscle tissues. However, the analysis of single myotubes is quite complicated because of the difficulty in distinguishing individual myotubes differentiated on a normal cell culture surface. In the present study, thin polydimethylsiloxane (PDMS) membranes, which have rectangular holes (30, 50, 100, and 200 microm in width; 500, 750, and 1000 microm in length) through them, were fabricated by using a photolithography technique and used for single myotube micropatterning. A bovine serum albumin-coated (BSA-coated) stencil membrane was placed on a cell culture surface and C2C12 myoblasts were seeded on it. Since the cells could not attach to the surface of the stencil membrane, the cell proliferated and differentiated into myotubes in the hole areas specifically. By peeling off the membrane, a micropattern of myotubes was obtained. It was revealed that the optimum width of rectangular holes for a micropattern of single myotubes was between 30 to 50 microm. Furthermore, by placing a membrane on a thermoresponsive culture surface, recovery of the micropatterned myotubes was possible by lowering the temperature. This method involving the stencil membranes and a thermoresponsive culture surface is useful for analyzing subcellular or single myotubes.

Scanning Electron Microscopy of the Egg of &lt;I&gt;Ochlerotatus albifasciatus&lt;/I&gt; (Diptera: Culicidae)

Ochlerotatus albifasciatus (Macquart) (Diptera: Culicidae) is a mosquito species of medical and veterinary importance, usually associated with temporary groundwater habitats. Eggs obtained from females collected in Córdoba city, in the center of Argentina, were studied using scanning electron microscopy. The eggs are elliptical in outline, measuring approximately 615.7 +/- 37.24 microm in length (1) and 180.7 +/- 22.87 microm in width (w), with an egg index (l/w ratio) of 2.9-3.7. The outer chorionic sculpture has racquet-shaped and wristwatch-shaped cells consisting of a small central tubercle surrounded by larger tubercles and followed by a row of smaller tubercles on one or both sides, respectively, although variations between regions of the egg were observed. The micropylar apparatus has a collar with evident molding and edges of determined length, albeit irregular, with defined margins for the transition area and a thickness of approximately 10.7 microm. The margins of the micropylar disc are raised and the disc measures approximately 21.1 microm in diameter. The micropyle is distinct.

Improved Hydrophobicity of Carbon Nanotube Arrays with Micropatterning

The hydrophobicity of vertically aligned multiwalled carbon nanotubes (MWCNTs) was improved through the creation of a parallel array of microwalls via a laser pruning technique. Changes to the hydrophobic nature of the patterned MWCNTs due to artificially induced roughness through variations in both the widths of the walls and the distance between adjacent walls, channel width, were investigated. The sample became more hydrophobic whenever water droplets landed on one microwall 7 or 13 microm in width. However, when a droplet bridged two microwalls, the surface became less hydrophobic. The optimal superhydrophobic MWCNT surface corresponded to a parallel array of microwalls with a width of 13 microm and a channel width of approximately 50 microm. Such findings could possibly serve as value-add for further developments in the creation of water-repelling CNT surfaces via micropatterning.

Thermus islandicus sp. nov., a mixotrophic sulfur-oxidizing bacterium isolated from the Torfajokull geothermal area

Strains PRI 2268 and PRI 3838(T) were isolated from two separate hot springs in the Torfajokull geothermal area of South Iceland. The cells were non-motile rods, approximately 0.3 microm in width and 1.5-2.5 microm in length. Electron microscopy revealed a Gram-negative cell-wall structure. The strains grew at 45-79 degrees C (optimum, 65 degrees C) and pH 5.5-10.5 (optimum, pH 6.0-7.0). 16S rRNA gene sequence analysis indicated that they formed a separate branch within the genus Thermus with 'Thermus kawarayensis' KW11 as their closest cultured relative (96.5 % similarity). The gene sequence similarities of both new isolates to Thermus aquaticus YT-1(T) and Thermus igniterrae RF-4(T) were 96.1 % and 95.5 %, respectively. DNA-DNA relatedness between strain PRI 3838(T) and 'T. kawarayensis' was 46.1 %. The DNA G+C content of strain PRI 3838(T) was 69.0 mol%. The predominant menaquinones, pigmentation, fatty acid profiles and phospholipid profiles of the novel strains were similar to those of other members of the genus Thermus. However, the new strains could be differentiated from the type strains of all other species of the genus Thermus by their lack of catalase activity and their utilization of only a few carbon sources. Furthermore, the novel strains exhibited mixotrophic growth with sulfur oxidation. On the basis of 16S rRNA gene sequence comparisons, DNA-DNA hybridization and physiological and biochemical characteristics, the new isolates represent a novel species. Since the species appears to be ubiquitous in Icelandic hot springs, the name Thermus islandicus sp. nov. is proposed. The type strain is PRI 3838(T) (=DSM 21543(T)=ATCC BAA-1677(T)).

Caldanaerovirga acetigignens gen. nov., sp. nov., an anaerobic xylanolytic, alkalithermophilic bacterium isolated from Trego Hot Spring, Nevada, USA

An anaerobic thermophilic bacterium, designated strain JW/SA-NV4(T), was isolated from a xylan-supplemented enrichment culture from Trego hot spring located within the Black Rock Desert (NV, USA). Cells were generally straight or slightly bent rod-shaped, 0.4-0.8 microm in width and 3-6 microm in length during exponential growth. Cells from stationary phase were variable in size and shape, showing curved or bent morphology. Motility was not seen and flagella were not observed in electron micrographs. Sporulation was not observed. Strain JW/SA-NV4(T) stained Gram-negative but is phylogenetically Gram-type positive. Growth occurred at pH(25 degrees C) 6.8-8.8, with optimum growth at pH 8.4; no growth occurred at pH 9.0 or above or at 6.5 or below. With glucose or xylose as the carbon source, strain JW/SA-NV4(T) grew at 44-74 degrees C; no growth occurred at 76 degrees C or above or at 42 degrees C or below. However, the optimum temperature was 62 and 66 degrees C when grown on glucose and xylose, respectively. The shortest doubling time observed with glucose was approximately 4 h, and with xylose approximately 3.4 h. Strain JW/SA-NV4(T) tolerated an atmosphere containing up to 0.1 % O(2); no growth occurred at a gas atmosphere of 0.2 % O(2). Chemo-organotrophic growth occurred with xylose, glucose, mannose, xylan, pyruvate, fructose, ribose, Casamino acids, mannitol, tryptone, peptone, cellobiose and yeast extract. When grown in mineral media containing 1 g yeast extract l(-1) as an electron donor, thiosulfate and sulfur were reduced to sulfide. The G+C content of the DNA was 38.6 mol% (HPLC). 16S rRNA gene sequence analysis placed strain JW/SA-NV4(T) within the order Thermoanaerobacterales and within the Thermoanaerobacterales Incertae Sedis Family III, specifically between taxa classified within the genera Thermosediminibacter and Thermovenabulum. The closest phylogenetic neighbours were Thermosediminibacter oceani JW/IW-1228P(T) (94.2 % 16S rRNA gene sequence similarity) and Thermosediminibacter litoriperuensis JW/YJL-1230-7/2(T) (94.0 %) [Lee, Y.-J., Wagner, I. D., Brice, M. E., Kevbrin, V. V., Mills, G. L., Romanek, C. S. & Wiegel, J. (2005). Extremophiles 9, 375-383]. Based on physiological and genotypic characteristics, strain JW/SA-NV4(T) (=DSM 18802(T)=ATCC BAA-1454(T)) is proposed to represent the type strain of a novel species in a novel genus, Caldanaerovirga acetigignens gen. nov., sp. nov.