Silicon consumption and population density changes of dominant planktonic diatoms in Lake Constance

(1) Interactions between silicon concentration and diatom growth have been studied in Lake Constance with data on number and volume of cells per unit volume of water and concentration of dissolved reactive silicon. (2) Concentration of dissolved reactive silicon was found to decrease markedly during the spring and summer growth period of diatoms in 1979 and 1980. (3) Amounts of silicon taken up per cell and per unit cell volume have been calculated from the changes in silicon concentration and the increases in population density for several diatom species. The silicon to cell quotient (mg per million cells) varied over a wide range from 0.005 for Stephanodiscus hantzschii to 0.72 for Synedra acus, while that of silicon per unit cell volume (mg mm-3) varied over a narrower range from 0. 10 for Stephanodiscus hantzschii and Fragilaria crotonesis to 0.36 for Synedra acus. (4) A four-compartment model, that allows estimates of the rates of growth and of loss of diatom populations to be calculated, is outlined. (5) Summer succession from Asterionellaformosa to either Fragilaria crotonensis (in 1979) or Stephanodiscus binderanus (in 1980) is explained by interspecific differences in the effects of silicon concentration on growth and death. For Asterionellaformosa, silicon depletion did not stop cell-division but led to the death of most of the population. For Fragilaria crotonensis and Stephanodiscus binderanus cell division stopped but cells did not die and population growth continued after the concentration of dissolved silicon had increased agai

Aestipascuomyces dupliciliberans gen. nov, sp. nov., the First Cultured Representative of the Uncultured SK4 Clade from Aoudad Sheep and Alpaca

We report on the isolation of the previously-uncultured Neocallimastigomycota SK4 lineage, by two independent research groups, from a wild aoudad sheep rumen sample (Texas, USA) and an alpaca fecal sample (Baden-Württemberg, Germany). Isolates from both locations showed near-identical morphological and microscopic features, forming medium-sized (2–5 mm) white filamentous colonies with a white center of sporangia, on agar roll tubes and a heavy biofilm in liquid media. Microscopic analysis revealed monocentric thalli, and spherical polyflagellated zoospores with 7–20 flagella. Zoospore release occurred through an apical pore as well as by sporangial wall rupturing, a duality that is unique amongst described anaerobic gut fungal strains. Isolates were capable of growing on a wide range of mono-, oligo-, and polysaccharide substrates as the sole carbon source. Phylogenetic assessment based on the D1–D2 28S large rRNA gene subunit (D1–D2 LSU) and internal transcribed spacer-1 (ITS-1) regions demonstrated high sequence identity (minimum identity of 99.07% and 96.96%, respectively) between all isolates; but low sequence identity (92.4% and 86.7%, respectively) to their closest cultured relatives. D1–D2 LSU phylogenetic trees grouped the isolates as a new monophyletic clade within the Orpinomyces–Neocallimastix–Pecoramyces–Feramyces–Ghazallamyces supragenus group. D1–D2 LSU and ITS-1 sequences recovered from the obtained isolates were either identical or displayed extremely high sequence similarity to sequences recovered from the same aoudad sheep sample on which isolation was conducted, as well as several sequences recovered from domestic sheep and few other herbivores. Interestingly, members of the SK4 clade seem to be encountered preferably in animals grazing on summer pasture. We hence propose accommodating these novel isolates in a new genus, Aestipascuomyces (derived from the Latin word for “summer pasture”), and a new species, A. dupliciliberans. The type strain is Aestipascuomycesdupliciliberans strain R4

Development and Evaluation of an Enzyme-Linked Immunosorbent Assay for Use in the Detection of Bovine Tuberculosis in Cattle

As a consequence of continued spillover of Mycobacterium bovis into cattle from wildlife reservoirs and increased globalization of cattle trade with associated transmission risks, new approaches such as vaccination and novel testing algorithms are seriously being considered by regulatory agencies for the control of bovine tuberculosis. Serologic tests offer opportunities for identification of M. bovis-infected animals not afforded by current diagnostic techniques. The present study describes assay development and field assessment of a new commercial enzyme-linked immunosorbent assay (ELISA) that detects antibody to M. bovis antigens MPB83 and MPB70 in infected cattle. Pertinent findings include the following: specific antibody responses were detected at 90 to 100 days after experimental M. bovis challenge, minimal cross-reactive responses were elicited by infection/sensitization with nontuberculous Mycobacterium spp., and the apparent sensitivity and specificity of the ELISA with naturally infected cattle were 63% and 98%, respectively, with sensitivity improv- ing as disease severity increased. The ELISA also detected infected animals missed by the routine tuberculin skin test, and antibody was detectable in bulk tank milk samples from M. bovis-infected dairy herds. A high-throughput ELISA could be adapted as a movement, border, or slaughter surveillance test, as well as a supplemental test to tuberculin skin testing

The periodicity of phytoplankton in Lake Constance (Bodensee) in comparison to other deep lakes of central Europe

Phytoplankton periodicity has been fairly regular during the years 1979 to 1982 in Lake Constance. Algal mass growth starts with the vernal onset of stratification; Cryptophyceae and small centric diatoms are the dominant algae of the spring bloom. In June grazing by zooplankton leads to a lsquoclear-water phasersquo dominated by Cryptophyceae. Algal summer growth starts under nutrient-saturated conditions with a dominance of Cryptomonas spp. and Pandorina morum. Depletion of soluble reactive phosphorus is followed by a dominance of pennate and filamentous centric diatoms, which are replaced by Ceratium hirundinella when dissolved silicate becomes depleted. Under calm conditions there is a diverse late-summer plankton dominated by Cyanophyceae and Dinobryon spp.; more turbulent conditions and silicon resupply enable a second summer diatom growth phase in August. The autumnal development leads from a Mougeotia — desmid assemblage to a diatom plankton in late autumn and winter. Inter-lake comparison of algal seasonality includes in ascending order of P-richness Königsee, Attersee, Walensee, Lake Lucerne, Lago Maggiore, Ammersee, Lake Zürich, Lake Geneva, Lake Constance. The oligotrophic lakes have one or two annual maxima of biomass; after the vernal maximum there is a slowly developing summer depression and sometimes a second maximum in autumn. The more eutrophic lakes have an additional maximum in summer. The number of floristically determined successional stages increases with increasing eutrophy, from three in Königsee and Attersee to eight in Lake Geneva and Lake Constance

Targeting Sphingosine Kinase 1 in Carcinoma Cells Decreases Proliferation and Survival by Compromising PKC Activity and Cytokinesis

Sphingosine kinases (SK) catalyze the phosphorylation of proapoptotic sphingosine to the prosurvival factor sphingosine 1-phosphate (S1P), thereby promoting oncogenic processes. Breast (MDA-MB-231), lung (NCI-H358), and colon (HCT 116) carcinoma cells were transduced with shRNA to downregulate SK-1 expression or treated with a pharmacologic SK-1 inhibitor. The effects of SK-1 targeting were investigated by measuring the level of intracellular sphingosine, the activity of protein kinase C (PKC) and cell cycle regulators, and the mitotic index. Functional assays included measurement of cell proliferation, colony formation, apoptosis, and cell cycle analysis. Downregulation of SK-1 or its pharmacologic inhibition increased intracellular sphingosine and decreased PKC activity as shown by reduced phosphorylation of PKC substrates. In MDA-MB-231 cells this effect was most pronounced and reduced cell proliferation and colony formation, which could be mimicked using exogenous sphingosine or the PKC inhibitor RO 31-8220. SK-1 downregulation in MDA-MB-231 cells increased the number of cells with 4N and 8N DNA content, and similar effects were observed upon treatment with sphingosine or inhibitors of SK-1 or PKC. Examination of cell cycle regulators unveiled decreased cdc2 activity and expression of Chk1, which may compromise spindle checkpoint function and cytokinesis. Indeed, SK-1 kd cells entered mitosis but failed to divide, and in the presence of taxol also failed to sustain mitotic arrest, resulting in further increased endoreduplication and apoptosis. Our findings delineate an intriguing link between SK-1, PKC and components of the cell cycle machinery, which underlines the significance of SK-1 as a target for cancer therapy

Genetic Loci Involved in Antibody Response to Mycobacterium avium ssp. paratuberculosis in Cattle

Background: Mycobacterium avium subsp. paratuberculosis (MAP) causes chronic enteritis in a wide range of animal species. In cattle, MAP causes a chronic disease called Johne's disease, or paratuberculosis, that is not treatable and the efficacy of vaccine control is controversial. The clinical phase of the disease is characterised by diarrhoea, weight loss, drop in milk production and eventually death. Susceptibility to MAP infection is heritable with heritability estimates ranging from 0.06 to 0.10. There have been several studies over the last few years that have identified genetic loci putatively associated with MAP susceptibility, however, with the availability of genome-wide high density SNP maker panels it is now possible to carry out association studies that have higher precision. Methodology/Principal Findings: The objective of the current study was to localize genes having an impact on Johne's disease susceptibility using the latest bovine genome information and a high density SNP panel (Illumina BovineSNP50 BeadChip) to perform a case/control, genome-wide association analysis. Samples from MAP case and negative controls were selected from field samples collected in 2007 and 2008 in the province of Lombardy, Italy. Cases were defined as animals serologically positive for MAP by ELISA. In total 966 samples were genotyped: 483 MAP ELISA positive and 483 ELISA negative. Samples were selected randomly among those collected from 119 farms which had at least one positive animal. Conclusion/Significance: The analysis of the genotype data identified several chromosomal regions associated with disease status: a region on chromosome 12 with high significance (P<5 710-6), while regions on chromosome 9, 11, and 12 had moderate significance (P<5 710-5). These results provide evidence for genetic loci involved in the humoral response to MAP. Knowledge of genetic variations related to susceptibility will facilitate the incorporation of this information into breeding programmes for the improvement of health status