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At least 37 records · Page 2Linked to original sources

Hydrogeology of the area near the J4 test cell, Arnold Air Force Base, Tennessee

The U.S. Air Force operates a major aerospace systems testing facility at Arnold Engineering Development Center (AEDC) in Coffee County, Tennessee. Dewatering operations at one of the test facilities, the J4 test cell, has affected the local ground-water hydrology. The J4 test cell is approximately 100 feet in diameter, extends approximately 250 feet below land surface, and penetrates several aquifers. Ground water is pumped continuously from around the test cell to keep the cell structurally intact. Because of the test cell's depth, dewatering has depressed water levels in the aquifers surrounding the site. The depressions that have developed exhibit anisotropy that is controlled by zones of high permeability in the aquifers. Additionally, contaminants - predominately volatile organic compounds - are present in the ground-water discharge from the test cell and in ground water at several other Installation Restoration Program (IRP) sites within the AEDC facility. The dewatering activities at J4 are drawing these contaminants from the nearby sites. The effects of dewatering at the J4 test cell were investigated by studying the lithologic and hydraulic characteristics of the aquifers, investigating the anisotropy and zones of secondary permeability using geophysical techniques, mapping the potentiometric surfaces of the underlying aquifers, and developing a conceptual model of the ground-water-flow system local to the test cell. Contour maps of the potentiometric surfaces in the shallow, Manchester, and Fort Payne aquifers (collectively, part of the Highland Rim aquifer system) show anisotropic water-level depressions centered on the J4 test cell. This anisotropy is the result of features of high permeability such as chert-gravel zones in the regolith and fractures, joints, and bedding planes in the bedrock. The presence of these features of high permeability in the Manchester aquifer results in complex flow patterns in the Highland Rim aquifers near the J4 test cell. The occurrence, distribution, and orientation of these features has a great effect on ground-water flow to the J4 test cell. The depression caused by dewatering extends out horizontally through the aquifers along the most permeable pathways. Since the aquifers above the Chattanooga Shale are not separated by distinct confining units, areas in adjacent aquifers above and below these zones of high permeability in the Manchester aquifer are also dewatered. Conditions in all Highland Rim aquifers approximate steady-state equilibrium because ground-water withdrawal at the test cell has been continuous since the late 1960's. The average ground-water discharge from the dewatering system at the J4 test cell was 105 gallons per minute, for 1992-95. The ground-water capture areas in each aquifer extend into all or parts of landfill #2 and leaching pit #2 (IRP site 1), the main testing area (IRP site 7), and the old fire training area (IRP site 10). IRP sites 8 and 12 are outside the ground-water capture areas. Of the 35 sampled wells in the J4 area, 10 produced water samples containing chlorinated organic compounds such as 1,2-dichloroethane (1,2-DCA), 1,1-dichloroethylene (1,1-DCE), and trichloroethylene (TCE) in concentrations which exceeded the Tennessee Department of Environment and Conservation Maximum Contaminant Levels (MCL's) for public water-supply systems. The highest concentrations were detected in samples from well AEDC-274 with 45 micrograms per liter (mg/L) 1,2-DCA, 320 mg/L 1,1-DCE, and 1,200 mg/L TCE. These compounds are synthetic and do not occur naturally in the environment. A sample of the ground-water discharge from the J4 test cell also contained concentrations of these compounds that exceed MCL's. Chlorinated organic compounds, including 1,2-DCA; 1,1-DCE; and TCE also have been detected at IRP sites 1, 7, 8, nd 10. The six dewatering wells surrounding the J4 test cell penetrate the Chattanooga Shale and are open to the Highland Rim aquifer system, there

Water-Resources Investigations Report↗

Functional classification of mitochondrion-rich cells in euryhaline Mozambique tilapia (Oreochromis mossambicus) embryos, by means of triple immunofluorescence staining for Na+/K+-ATPase, Na +/K+/2Cl- cotransporter and CFTR anion channel

Mozambique tilapia Oreochromis mossambicus embryos were transferred from freshwater to seawater and vice versa, and short-term changes in the localization of three major ion transport proteins, Na+/K +-ATPase, Na+/K+/2Cl- cotransporter (NKCC) and cystic fibrosis transmembrane conductance regulator (CFTR) were examined within mitochondrion-rich cells (MRCs) in the embryonic yolk-sac membrane. Triple-color immunofluorescence staining allowed us to classify MRCs into four types: type I, showing only basolateral Na+/K +-ATPase staining; type II, basolateral Na+/K +-ATPase and apical NKCC; type III, basolateral Na+/K +-ATPase and basolateral NKCC; type IV, basolateral Na +/K+-ATPase, basolateral NKCC and apical CFTR. In freshwater, type-I, type-II and type-III cells were observed. Following transfer from freshwater to seawater, type-IV cells appeared at 12 h and showed a remarkable increase in number between 24 h and 48 h, whereas type-III cells disappeared. When transferred from seawater back to freshwater, type-IV cells decreased and disappeared at 48 h, type-III cells increased, and type-II cells, which were not found in seawater, appeared at 12 h and increased in number thereafter. Type-I cells existed consistently irrespective of salinity changes. These results suggest that type I is an immature MRC, type II is a freshwater-type ion absorptive cell, type III is a dormant type-IV cell and/or an ion absorptive cell (with a different mechanism from type II), and type IV is a seawater-type ion secretory cell. The intracellular localization of the three ion transport proteins in type-IV cells is completely consistent with a widely accepted model for ion secretion by MRCs. A new model for ion absorption is proposed based on type-II cells possessing apical NKCC.

Journal of Experimental Biology↗

An assessment of the effects of cell size on AGNPS modeling of watershed runoff

This study investigates the changes in simulated watershed runoff from the Agricultural NonPoint Source (AGNPS) pollution model as a function of model input cell size resolution for eight different cell sizes (30 m, 60 m, 120 m, 210 m, 240 m, 480 m, 960 m, and 1920 m) for the Little River Watershed (Georgia, USA). Overland cell runoff (area-weighted cell runoff), total runoff volume, clustering statistics, and hot spot patterns were examined for the different cell sizes and trends identified. Total runoff volumes decreased with increasing cell size. Using data sets of 210-m cell size or smaller in conjunction with a representative watershed boundary allows one to model the runoff volumes within 0.2 percent accuracy. The runoff clustering statistics decrease with increasing cell size; a cell size of 960 m or smaller is necessary to indicate significant high-runoff clustering. Runoff hot spot areas have a decreasing trend with increasing cell size; a cell size of 240 m or smaller is required to detect important hot spots. Conclusions regarding cell size effects on runoff estimation cannot be applied to local watershed areas due to the inconsistent changes of runoff volume with cell size; but, optimal cells sizes for clustering and hot spot analyses are applicable to local watershed areas due to the consistent trends.

Cartography and Geographic Information Science↗

Establishment of a cell line with reticulo-endothelial characteristics from a rainbow trout spleen explant

A cell line from rainbow trout ( Oncorhynchus mykiss ) with phagocytic-like characteristics was established. First we found that it was necessary to inject the fish with antibiotics (penicillinstreptomycin) in order to clear tissue of bacterial contamination. Two days later, the spleen was excised and minced with scissors in MEM10. The large number of erythrocytes in the suspension were removed by low centrifugation, then the tissue fragments were incubated on tissue culture flasks at 18°C for 30 days with complete changes of media every other day. After 30 days in culture, primary explants from spleen were composed of three types of cells. The majority were large mononucleate, plecmorphic cells with thinly spread cytoplasm. These cells were trypsinized and subcultured. Another cell types was simple, round cells, somewhat similar to mature lymphocytes, while the third type appeared to be similar to fibroblastic cells. After 9 months with the culture in 17 to 20 passages, the predominate cell type was showing similarities to phagocytic cell. These cells had a marked reticular cytoplasm and grew individually without parallel orientations. By enzyme-specific stains, the cells were observed to have abundant acid phosphatase and β-glucuronidase activities of the cytoplasm. They were negative for peroxidase. The cells were also found to have highly adherent and phagocytic properties when incubated with latex particles. The morphological, enzymatic, and functional findings suggested the possibility that these cells in culture have some properties similar to the cells which are composed of reticulo-endothelial system.

Fish Pathology↗

Fish T cells: recent advances through genomics

This brief review is intended to provide a concise overview of the current literature concerning T cells, advances in identifying distinct T cell functional subsets, and in distinguishing effector cells from memory cells. We compare and contrast a wealth of recent progress made in T cell immunology of teleost, elasmobranch, and agnathan fish, to knowledge derived from mammalian T cell studies. From genome studies, fish clearly have most components associated with T cell function and we can speculate on the presence of putative T cell subsets, and the ability to detect their differentiation to form memory cells. Some recombinant proteins for T cell associated cytokines and antibodies for T cell surface receptors have been generated that will facilitate studying the functional roles of teleost T cells during immune responses. Although there is still a long way to go, major advances have occurred in recent years for investigating T cell responses, thus phenotypic and functional characterization is on the near horizon.

Developmental and Comparative Immunology↗

Temperature comparisons for antibody production in vitro by plaque-forming cells from trout Salmo gairdneri (Richardson), and mice

Anterior kidney and splenic cells were taken from rainbow trout and splenic cells from BALB/c mice immunized with a T-dependent (sheep red blood cells) or T-independent (DNP-Ficoll) antigen. The cells were incubated at different temperatures in Jerne plaque assays (direct or passive haemolytic plaque assays). The optimum numbers of in vitro plaque-forming cells (PFC) after incubation with homologous complement were directly correlated with normal body temperatures of the respective species. The optimum incubation temperature was 37°C for mouse cells and 10°C for fish cells. Incubation of mouse cells at lower temperatures of 30, 20, 10, 4 or 0°C appeared to yield a direct line reduction in numbers of PFC. Trout cells developed significantly fewer PFC at 4 and 20°C and none at 30°C or above; however, significant numbers still appeared at 0°C. More PFC per million white blood cells were obtained from the anterior kidney; however, related to temperatures, no differences in development of numbers of PFC could be seen between the spleen and anterior kidney cells of trout. When the incubation time was lengthened for both trout and mouse cells held at low temperatures, the numbers of PFC approached those of the cells incubated at the optimum temperatures for 10 h.

Journal of Fish Biology↗

Why large cells dominate estuarine phytoplankton

Surveys across the world oceans have shown that phytoplankton biomass and production are dominated by small cells (picoplankton) where nutrient concentrations are low, but large cells (microplankton) dominate when nutrient-rich deep water is mixed to the surface. I analyzed phytoplankton size structure in samples collected over 25 yr in San Francisco Bay, a nutrient-rich estuary. Biomass was dominated by large cells because their biomass selectively grew during blooms. Large-cell dominance appears to be a characteristic of ecosystems at the land–sea interface, and these places may therefore function as analogs to oceanic upwelling systems. Simulations with a size-structured NPZ model showed that runs of positive net growth rate persisted long enough for biomass of large, but not small, cells to accumulate. Model experiments showed that small cells would dominate in the absence of grazing, at lower nutrient concentrations, and at elevated (+5°C) temperatures. Underlying these results are two fundamental scaling laws: (1) large cells are grazed more slowly than small cells, and (2) grazing rate increases with temperature faster than growth rate. The model experiments suggest testable hypotheses about phytoplankton size structure at the land–sea interface: (1) anthropogenic nutrient enrichment increases cell size; (2) this response varies with temperature and only occurs at mid-high latitudes; (3) large-cell blooms can only develop when temperature is below a critical value, around 15°C; (4) cell size diminishes along temperature gradients from high to low latitudes; and (5) large-cell blooms will diminish or disappear where planetary warming increases temperature beyond their critical threshold.

Limnology and Oceanography↗

Calcein AM release-based cytotoxic cell assay for fish leucocytes

A non-specific cytotoxic cell assay for fish is presented that is based on the release of the activated fluorochrome calcein AM from lysed carp epithelioma papulosum cyprini (EPC) cells. To establish the suitability of treating EPC cells with calcein AM the uptake and spontaneous release of the calcein AM by the EPC cells was evaluated. Incubation of 5 μM calcein AM in culture medium with 1×10 5 EPC cells well −1 for a minimum of 3 h provided sufficient labelling. Spontaneous release of fluorescence from the labelled EPC cells during 10 h of post labelling incubation ranged from 30 to 39% of the total observed fluorescence. Cytotoxic activity of trout leucocytes was evaluated at three leucocyte to target cell ratios (10:1, 2:1 and 1:1) following incubation (4, 6, 8, and 10 h) with calcein AM-labelled EPC cells at 15 °C. In some instances, the monoclonal antibody specific for the NCC surface receptor NCCRP-1 (MAb5C.6) was included in the cultures. The activity of NCC cells was significantly inhibited in the presence of 0.25 μg well −1 of MAb5C.6 relative to no antibody ( P ≤0.013) or an equal amount of an unrelated antibody ( P ≤0.001). Average maximum observed percent cytotoxic cell activity of ∼18% was observed following 8 h of incubation at the 2:1 and 1:1 leucocyte to target cell ratios. Percent cytotoxic cell activity using calcein AM was similar to values reported for rainbow trout leucocytes using the 51 Cr-release assay.

Fish and Shellfish Immunology↗

In vitro biology of fibropapilloma-associated turtle herpesvirus and host cells in Hawaiian green turtles (Chelonia mydas)

Fibropapillomatosis (FP) of green turtles has a global distribution and causes debilitating tumours of the skin and internal organs in several species of marine turtles. FP is associated with a presently non-cultivable alphaherpesvirus Chelonid fibropapilloma-associated herpesvirus (CFPHV). Our aims were to employ quantitative PCR targeted to pol DNA of CFPHV to determine (i) if DNA sequesters by tumour size and/or cell type, (ii) whether subculturing of cells is a viable strategy for isolating CFPHV and (iii) whether CFPHV can be induced to a lytic growth cycle in vitro using chemical modulators of replication (CMRs), temperature variation or co-cultivation. Additional objectives included determining whether non-tumour and tumour cells behave differently in vitro and confirming the phenotype of cultured cells using cell-type-specific antigens. CFPHV pol DNA was preferentially concentrated in dermal fibroblasts of skin tumours and the amount of viral DNA per cell was independent of tumour size. Copy number of CFPHV pol DNA per cell rapidly decreased with cell doubling of tumour-derived fibroblasts in culture. Attempts to induce viral replication in known CFPHV-DNA-positive cells using temperature or CMR failed. No significant differences were seen in in vitro morphology or growth characteristics of fibroblasts from tumour cells and paired normal skin, nor from CFPHV pol-DNA-positive intestinal tumour cells. Tumour cells were confirmed as fibroblasts or keratinocytes by positive staining with anti-vimentin and anti-pancytokeratin antibodies, respectively. CFPHV continues to be refractory to in vitro cultivation.

Hawai'i↗

The impact of metabolic state on Cd adsorption onto bacterial cells

This study examines the effect of bacterial metabolism on the adsorption of Cd onto Gram-positive and Gram-negative bacterial cells. Metabolically active Gram-positive cells adsorbed significantly less Cd than non-metabolizing cells. Gram-negative cells, however, showed no systematic difference in Cd adsorption between metabolizing and non-metabolizing cells. The effect of metabolism on Cd adsorption to Gram-positive cells was likely due to an influx of protons in and around the cell wall from the metabolic proton motive force, promoting competition between Cd and protons for adsorption sites on the cell wall. The relative lack of a metabolic effect on Cd adsorption onto Gram-negative compared to Gram-positive cells suggests that Cd binding in Gram-negative cells is focused in a region of the cell wall that is not reached, or is unaffected by this proton flux. Thermodynamic modeling was used to estimate that proton pumping causes the pH in the cell wall of metabolizing Gram-positive bacteria to decrease from the bulk solution value of 7.0 to approximately 5.7.

Geobiology↗

Recycled Cell Phones - A Treasure Trove of Valuable Metals

This U.S. Geological Survey (USGS) Fact Sheet examines the potential value of recycling the metals found in obsolete cell phones. Cell phones seem ubiquitous in the United States and commonplace throughout most of the world. There were approximately 1 billion cell phones in use worldwide in 2002. In the United States, the number of cell phone subscribers increased from 340,000 in 1985 to 180 million in 2004. Worldwide, cell phone sales have increased from slightly more than 100 million units per year in 1997 to an estimated 779 million units per year in 2005. Cell phone sales are projected to exceed 1 billion units per year in 2009, with an estimated 2.6 billion cell phones in use by the end of that year. The U.S. Environmental Protection Agency estimated that, by 2005, as many as 130 million cell phones would be retired annually in the United States. The nonprofit organization INFORM, Inc., anticipated that, by 2005, a total of 500 million obsolete cell phones would have accumulated in consumers' desk drawers, store rooms, or other storage, awaiting disposal. Typically, cell phones are used for only 1 1/2 years before being replaced. Less than 1 percent of the millions of cell phones retired and discarded annually are recycled. When large numbers of cell phones become obsolete, large quantities of valuable metals end up either in storage or in landfills. The amount of metals potentially recoverable would make a significant addition to total metals recovered from recycling in the United States and would supplement virgin metals derived from mining.

Fact Sheet↗

Assessment of significant sand resources in Federal and California State Waters of the San Francisco, Oceanside, and Silver Strand littoral cell study areas along the continental shelf of California

Executive Summary The Sand Resources Project was established through collaborative agreements between the U.S. Geological Survey (USGS), the Bureau of Ocean Energy Management (BOEM), and the California Ocean Protection Council (OPC) with the purpose of evaluating sand and gravel resources in Federal and California State Waters for potential use in future beach-nourishment projects. Project partners worked in collaboration with California Coastal Sediment Management Workgroup (CSMW) members to define priority study areas for this work based on the potential for finding sand within the broader region and the needs for this sand as shown by beach erosion areas of concern in the adjacent littoral cells. The final study areas were defined to be (1) the San Francisco Littoral Cell, (2) the Oceanside Littoral Cell, and (3) the Silver Strand Littoral Cell. A two-stage approach was used to assess the study areas. The initial stage was a synthesis of the existing geophysical and sediment-sampling data in each area. This allowed for evaluations of the data availability, data gaps, and general patterns of sediment thickness and grain size. This synthesis was published in a separate USGS open-file report (Warrick and others, 2022). The findings from this assessment were used to refine study area boundaries and develop sampling plans for stage two of the project. Stage two of the project is the collection, processing, and synthesis of new data, including high-resolution geophysical surveys and sediment cores—this report addresses the second stage. The work focuses on two of the study areas—the San Francisco and the Oceanside Littoral Cells, where several research cruises have been conducted. A more limited, exploratory approach was used for the Silver Strand Littoral Cell, owing to the lack of existing high-resolution bathymetric data for this study area. The data collected provide new information about the three study areas, including sediment thickness, grain-size distributions, and total organic carbon. Sediment in all three study areas of the Sand Resources Study was suitable for beach nourishment, as reflected by their grain-size distributions and sediment thicknesses. For example, sandy sediment in the San Francisco Littoral Cell study area was on and immediately outside of the ebb-tidal bar of the San Francisco Bay, a landform that has a strong influence on grain-size patterns of the region. The presence of thick sediment deposits in this area was interpreted to be a function of tectonics, which has caused physical features that include a graben north of the Golden Gate whose deposits were thicker and siltier than the remaining area. Sandy sediment on the inner and outer parts of the continental shelf in the Oceanside Littoral Cell may be useful for nourishment, whereas the midshelf between these areas was dominated by silty sediment. Sediment in the Silver Strand Littoral Cell, which was only sampled selectively, had the greatest potential for beach nourishment because of the greater prevalence of beach-comparable grain sizes, especially in the more distal and deeper areas where medium sands were found. The Sand Resources Project did identify several sandy regions of the continental shelf that are deeper than dredging technologies currently (2022) available in the United States, which are generally limited to 30 meters (m) water depth or less. Although sandy sediment exists in all three study areas at water depths of 30 m or less, additional sediment supplies—most of which are in Federal Waters—are present in deeper settings, especially for the Oceanside and Silver Strand Littoral Cell study areas. Although the Silver Strand Littoral Cell study area was found to be considerably replete in sand resources, these conclusions are based on a limited sampling exercise across that study area. Thus, it may be beneficial to complete a more thorough characterization of the sediment resources in the Silver Strand Littoral Cell study area if it is determined that a need for sandy coastal sediment exists in this region. As a result of the Sand Resources Project, several areas of sand resources in Federal and California State Waters were found where they were previously unknown. As such, this project may provide important data for future coastal-management decisions in California, and it should provide a model for future investigations of sediment resources in other regions of the State.

California↗

Assessing humoral and cell-mediated immune response in Hawaiian green turtles, Chelonia mydas

Seven immature green turtles, Chelonia mydas , captured from Kaneohe Bay on the island of Oahu were used to evaluate methods for assessing their immune response. Two turtles each were immunized intramuscularly with egg white lysozyme (EWL) in Freund’s complete adjuvant, Gerbu, or ISA-70; a seventh turtle was immunized with saline only and served as a control. Humoral immune response was measured with an indirect enzyme linked immunosorbent assay (ELISA). Cell-mediated immune response was measured using in vitro cell proliferation assays (CPA) using whole blood or peripheral blood mononuclear cells (PBM) cultured with concanavalin A (ConA), phytohaemagglutinin (PHA), or soluble egg EWL antigen. All turtles, except for one immunized with Gerbu and the control, produced a detectable humoral immune response by 6 weeks which persisted for at least 14 weeks after a single immunization. All turtles produced an anamnestic humoral immune response after secondary immunization. Antigen specific cell-mediated immune response in PBM was seen in all turtles either after primary or secondary immunization, but it was not as consistent as humoral immune response; antigen specific cell-mediated immune response in whole blood was rarely seen. Mononuclear cells had significantly higher stimulation indices than whole blood regardless of adjuvant, however, results with whole blood had lower variability. Both Gerbu and ISA-70 appeared to potentiate the cell-mediated immune response when PBM or whole blood were cultured with PHA. This is the first time cell proliferation assays have been compared between whole blood and PBM for reptiles. This is also the first demonstration of antigen specific cell-mediated response in reptiles. Cell proliferation assays allowed us to evaluate the cell-mediated immune response of green turtles. However, CPA may be less reliable than ELISA for detecting antigen specific immune response. Either of the three adjuvants appears suitable to safely elicit a detectable immune response in green turtles.

Hawai'i↗

Evolution of a minimal cell

Possessing only essential genes, a minimal cell can reveal mechanisms and processes that are critical for the persistence and stability of life 1,2 . Here we report on how an engineered minimal cell 3,4 contends with the forces of evolution compared with the Mycoplasma mycoides non-minimal cell from which it was synthetically derived. Mutation rates were the highest among all reported bacteria, but were not affected by genome minimization. Genome streamlining was costly, leading to a decrease in fitness of greater than 50%, but this deficit was regained during 2,000 generations of evolution. Despite selection acting on distinct genetic targets, increases in the maximum growth rate of the synthetic cells were comparable. Moreover, when performance was assessed by relative fitness, the minimal cell evolved 39% faster than the non-minimal cell. The only apparent constraint involved the evolution of cell size. The size of the non-minimal cell increased by 80%, whereas the minimal cell remained the same. This pattern reflected epistatic effects of mutations in ftsZ , which encodes a tubulin-homologue protein that regulates cell division and morphology 5,6 . Our findings demonstrate that natural selection can rapidly increase the fitness of one of the simplest autonomously growing organisms. Understanding how species with small genomes overcome evolutionary challenges provides critical insights into the persistence of host-associated endosymbionts, the stability of streamlined chassis for biotechnology and the targeted refinement of synthetically engineered cells 2,7,8,9 .

Nature↗

Simultaneous reduction of nitrate and selenate by cell suspensions of selenium-respiring bacteria

Washed-cell suspensions of Sulfurospirillum barnesii reduced selenate [Se(VI)] when cells were cultured with nitrate, thiosulfate, arsenate, or fumarate as the electron acceptor. When the concentration of the electron donor was limiting, Se(VI) reduction in whole cells was approximately fourfold greater in Se(VI)-grown cells than was observed in nitrate-grown cells; correspondingly, nitrate reduction was ∼11-fold higher in nitrate-grown cells than in Se(VI)-grown cells. However, a simultaneous reduction of nitrate and Se(VI) was observed in both cases. At nonlimiting electron donor concentrations, nitrate-grown cells suspended with equimolar nitrate and selenate achieved a complete reductive removal of nitrogen and selenium oxyanions, with the bulk of nitrate reduction preceding that of selenate reduction. Chloramphenicol did not inhibit these reductions. The Se(VI)-respiring haloalkaliphile Bacillus arsenicoselenatis gave similar results, but its Se(VI) reductase was not constitutive in nitrate-grown cells. No reduction of Se(VI) was noted for Bacillus selenitireducens , which respires selenite. The results of kinetic experiments with cell membrane preparations of S. barnesii suggest the presence of constitutive selenate and nitrate reduction, as well as an inducible, high-affinity nitrate reductase in nitrate-grown cells which also has a low affinity for selenate. The simultaneous reduction of micromolar Se(VI) in the presence of millimolar nitrate indicates that these organisms may have a functional use in bioremediating nitrate-rich, seleniferous agricultural wastewaters. Results with 75 Se-selenate tracer show that these organisms can lower ambient Se(VI) concentrations to levels in compliance with new regulations proposed for release of selenium oxyanions into the environment.

California↗

Influence of salinity on the localization of Na+/K +-ATPase, Na+/K+/2Cl- cotransporter (NKCC) and CFTR anion channel in chloride cells of the Hawaiian goby (Stenogobius hawaiiensis)

Na+/K+-ATPase, Na+/K+/2Cl- cotransporter (NKCC) and cystic fibrosis transmembrane conductance regulator (CFTR) are the three major transport proteins thought to be involved in chloride secretion in teleost fish. If this is the case, the levels of these transporters should be high in chloride cells of seawater-acclimated fish. We therefore examined the influence of salinity on immunolocalization of Na +/K+-ATPase, NKCC and CFTR in the gills of the Hawaiian goby (Stenogobius hawaiiensis). Fish were acclimated to freshwater and 20??? and 30??? seawater for 10 days. Na+/K +-ATPase and NKCC were localized specifically to chloride cells and stained throughout most of the cell except for the nucleus and the most apical region, indicating a basolateral/tubular distribution. All Na+/K +-ATPase-positive chloride cells were also positive for NKCC in all salinities. Salinity caused a slight increase in chloride cell number and size and a slight decrease in staining intensity for Na+/K +-ATPase and NKCC, but the basic pattern of localization was not altered. Gill Na+/K+-ATPase activity was also not affected by salinity. CFTR was localized to the apical surface of chloride cells, and only cells staining positive for Na+/K+-ATPase were CFTR-positive. CFTR-positive cells greatly increased in number (5-fold), area stained (53%) and intensity (29%) after seawater acclimation. In freshwater, CFTR immunoreactivity was light and occurred over a broad apical surface on chloride cells, whereas in seawater there was intense immunoreactivity around the apical pit (which was often punctate in appearance) and a light subapical staining. The results indicate that Na+/K +-ATPase, NKCC and CFTR are all present in chloride cells and support current models that all three are responsible for chloride secretion by chloride cells of teleost fish.

Journal of Experimental Biology↗

Cloned embryos from semen. Part 2: Intergeneric nuclear transfer of semen-derived eland (Taurotragus oryx) epithelial cells into bovine oocytes

The production of cloned offspring by nuclear transfer (NT) of semen-derived somatic cells holds considerable potential for the incorporation of novel genes into endangered species populations. Because oocytes from endangered species are scarce, domestic species oocytes are often used as cytoplasts for interspecies NT. In the present study, epithelial cells isolated from eland semen were used for intergeneric transfer (IgNT) into enucleated bovine oocytes and compared with bovine NT embryos. Cleavage rates of bovine NT and eland IgNT embryos were similar (80 vs. 83%, respectively; p > 0.05); however, development to the morula and blastocyst stage was higher for bovine NT embryos (38 and 21%, respectively; p < 0.0001), than for eland IgNT embryos (0.5 and 0%, respectively). DNA synthesis was not observed in either bovine NT or eland IgNT cybrids before activation, but in 75 and 70% of bovine NT and eland igNT embryos, respectively, cell-cycle resumption was observed at 16 h postactivation (hpa). For eland IgNT embryos, 13% had ???8 cells at 84 hpa, while 32% of the bovine NT embryos had ???8 cells at the same interval. However, 100 and 66% of bovine NT and eland IgNT embryos, respectively, that had ???8 cells synthesized DNA. From these results we concluded that (1) semen-derived epithelial cell nuclei can interact and be transcriptionally controlled by bovine cytoplast, (2) the first cell-cycle occurred in IgNT embryos, (3) a high frequency of developmental arrest occurs before the eight-cell stage in IgNT embryos, and (4) IgNT embryos that progress through the early cleavage stage arrest can (a) synthesize DNA, (b) progress through subsequent cell cycles, and (c) may have the potential to develop further. ?? 2008 Mary Ann Liebert, Inc.

Cloning and Stem Cells↗

Hydraulic characteristics of a rectangular mixed-cell rearing unit

We describe a fish rearing unit modification that establishes mixed flow reactor (MFR) behavior in a rectangular vessel so as to eliminate metabolite concentration gradients, increase current velocities and improve solids scour at low water exchange rates. A standard raceway section 14.5 m long was modified to create six counter-rotating mixed cells, each 2.4 m wide by 2.4 m long. Cells receive water from vertical pipe sections extending to the tank floor and positioned in the corners of the cells. Vertical pipe sections were fabricated with jet ports that direct water into the cells tangentially to establish rotary circulation. Water exits each cell through a centrally located floor drain. Hydraulic characteristics of the tank are described based on an analysis of residence time distribution, current velocities and cell-to-cell water exchange rates. The latter was calculated based on a proposed hydraulics model and tracer concentrations established at a steady state. Vessel dispersion numbers measured with water exchange rates of 1.0 and 1.3 per hour, indicate tank performance approximates that of a MFR with the active tank volume representing 77.9–100% of total tank volume. Water velocities measured at 216 stations averaged 0.14, 0.12 and 0.12 m/s for the tank surface, mid-depth and near bottom regions. The benthic shear stress calculated given surface velocity was 0.04 N/m 2 . This was sufficient to scour and purge fecal solids produced by 2010 kg of lake trout that averaged 44.3 cm in total length. Cell interaction was significant with cell-to-cell exchange rates averaging 3.9 times the tank inflow rate. Energy requirements of the rearing unit were kept low (1.32 m water gauge pressure; 6.7 W/m 3 ) through use of a large number of low velocity jets.

Aquacultural Engineering↗