Crop uptake and leaching of 15N applied in ruminant slurry with selectively labelled faeces and urine fractions

Two animal slurries either labelled with ¹⁵N in the urine or in the faeces fraction, were produced by feeding a sheep with unlabelled and ¹⁵N-labelled hay and collecting faeces and urine separately. The slurries were applied (12 g total N ^-2) to a coarse sand and a sandy loam soil confined in lysimeters and growing spring barley (Hordeum vulgare L). Reference lysimeters without slurry were supplied with¹⁵ NH₄ ¹⁵NO³ corresponding to the inorganic N applied with the slurries (6 g N m^-2). In the second year, all lysimeters received unlabelled mineral fertilizer (6 g N m^-2) and grew spring barley. N harvested in the two crops (grain + straw) and the loss of nitrate by leaching were determined. ¹⁵N in the urine fraction was less available for crop uptake than mineral fertilizer ¹⁵N. The first barley crop on the sandy loam removed 49% of the ¹⁵N applied in mineral fertilizer and 36% of that applied with urine. The availability of fertilizer ¹⁵N (36%) and urine¹⁵ N (32%) differed less on the coarse sand. Of the¹⁵ N added with the faeces fraction, 12–14% was taken up by the barley crop on the two soils. N mineralized from faeces compensated for the reduced availability of urine N providing a similar or higher crop N uptake in manured lysimeters compared with mineral fertilized ones.About half of the total N uptake in the first crop originated from the N applied either as slurry or mineral fertilizer. The remaining N was derived from the soil N pool. Substantially smaller but similar proportions of¹⁵ N from faeces, urine and fertilizer were found in the second crop. The similar recoveries indicated a slow mineralization rate of the residual faeces N since more faeces was left in the soil after the first crop.More N was lost by leaching from manured lysimeters but as a percentage of N applied, losses were similar to those from mineral fertilizer. During the first and second winter, 3–5% and 1–3%, respectively, of the ¹⁵N in slurry and mineral fertilizer was leached as nitrate. Thus slurry N applied in spring just before sowing did not appear to be more prone to loss by nitrate leaching than N given in mineral fertilizer. Slurry N accounted for a higher proportion of the N leached, however, because more N was added in this treatment.     

Numerical taxonomic study of thermophilic Bacillus isolated from three geographically separated deep-sea hydrothermal vents

Abstract: A numerical taxonomic study has been carried out with 80 strains, newly isolated, from three geographically separated deep-sea hydrothermal vents (Mid-Atlantic Ridge, Guaymas Basin and Lau Basin) and eleven thermophilic reference strains representing 11 Bacillus species. The deep-sea isolates were all halotolerant spore-forming rods and grew aerobically above 65°C. Results from unweighted average linkage cluster analysis of a similarity matrix derived from the simple matching coefficient, showed formation of nine major phena, which were defined at the 83% similarity level or above. Seven phena were composed exclusively of strains isolated from the same site (4 from Mid-Atlantic Ridge, 1 from Guaymas Basin and 2 from Lau Basin). The majority of the Lau Basin isolates clustered with 6 of the reference strains in one phenon, while isolates from Mid-Atlantic Ridge and Guaymas Basin were found separated from this phenon at the 69% similarity level. The other reference strains showed less than 69% similarity with the deep-sea isolates.     

Bacterial composition and succession during storage of North-Atlantic cod (Gadus morhua) at superchilled temperatures

Background  

The bacteriology during storage of the North-Atlantic cod has been investigated for the past decades using conventional cultivation strategies which have generated large amount of information. This paper presents a study where both conventional cultivation and cultivation independent approaches were used to investigate the bacterial succession during storage of cod loins at chilled and superchilled temperatures.     

Actinobacteria—An Ancient Phylum Active in Volcanic Rock Weathering

A molecular biological analysis of Icelandic volcanic rocks of different compositions and glassiness revealed the presence of Actinobacteria as an abundant phylum. In outcrops of basaltic glass they were the dominant bacterial phylum. A diversity of Actinobacteria were cultured from the rocks on rock-agar plates showing that they are capable of growing on rock-derived nutrient sources and that many of the taxa identified by molecular methods are viable, potentially active members of the community. Laboratory batch-culture experiments using a Streptomyces isolate showed that it was capable of enhancing the release of major elements from volcanic rocks, including weathered basaltic glass, crystalline basalt and komatiite, when provided with a carbon source. Actinobacteria of a variety of other sub-orders were also capable of enhancing volcanic rock weathering, measured as Si release. However, most strains did not significantly increase the weathering of the silica-rich rock, obsidian. These data show that Actinobacteria can contribute to volcanic rock weathering and, therefore, the carbonate-silicate cycle. Given their ancient lineage, it is likely they have played a role in rock weathering for over two billion years.     

The Toll-like receptor 7/8 agonist resiquimod greatly increases the immunostimulatory capacity of human acute myeloid leukemia cells

Immunotherapy for leukemia is a promising targeted strategy to eradicate residual leukemic cells after standard therapy, in order to prevent relapse and to prolong the survival of leukemia patients. However, effective anti-leukemia immune responses are hampered by the weak immunogenicity of leukemic cells. Therefore, much effort is made to identify agents that could increase the immunogenicity of leukemic cells and activate the immune system. Synthetic agonists of Toll-like receptor (TLR)7 and TLR8 are already in use as anticancer treatment, because of their ability to activate several immune pathways simultaneously, resulting in effective antitumor immunity. However, for leukemic cells little is known about the expression of TLR7/8 and the direct effects of their agonists. We hypothesized that TLR7/8 agonist treatment of human acute myeloid leukemia (AML) cells would lead to an increased immunogenicity of AML cells. We observed expression of TLR7 and TLR8 in primary human AML cells and AML cell lines. Passive pulsing of primary AML cells with the TLR7/8 agonist R-848 resulted in increased expression of MHC molecules, production of proinflammatory cytokines, and enhanced allogeneic naïve T cell-stimulatory capacity. These effects were absent or suboptimal if R-848 was administered intracellularly by electroporation. Furthermore, when AML cells were cocultured with allogeneic PBMC in the presence of R-848, interferon (IFN)-γ was produced by allogeneic NK and NKT cells and AML cells were killed. In conclusion, the immunostimulatory effect of the TLR7/8 agonist R-848 on human AML cells could prove useful for the design of TLR-based immunotherapy for leukemia.     

Investigation of the Microbial Ecology of Intertidal Hot Springs by Using Diversity Analysis of 16S rRNA and Chitinase Genes

The microbial diversity of intertidal hot springs on the seashore of northwest Iceland was examined by combining directed in situ enrichments, artificial support colonization, and mat sampling. Analysis of 16S rRNA genes revealed the presence of clones related to both marine and terrestrial, thermophilic, mesophilic, and psychrophilic microorganisms scattered among 11 bacterial divisions. No archaea were found. The species composition of the enrichments was affected by the length of the hot periods experienced at low tide and was very different from those found in the biomass. A total of 36 chitinase genes were detected by molecular screening of the samples with degenerate primers for glycoside hydrolase family 18. The chitinase gene diversity was at least twofold higher in the enrichment samples than in the controls, indicating that a much higher diversity of hydrolytic genes can be accessed with this approach.     

Identification of RAS-mitogen-activated protein kinase signaling pathway modulators in an ERF1 redistribution screen

The RAS-mitogen-activated protein kinase (MAPK) signaling pathway has a central role in regulating the proliferation and survival of both normal and tumor cells. This pathway has been 1 focus area for the development of anticancer drugs, resulting in several compounds, primarily kinase inhibitors, in clinical testing. The authors have undertaken a cell-based, high-throughput screen using a novel ERF1 Redistribution assay to identify compounds that modulate the signaling pathway. The hit compounds were subsequently tested for activity in a functional cell proliferation assay designed to selectively detect compounds inhibiting the proliferation of MAPK pathway-dependent cancer cells. The authors report the identification of 2 cell membrane-permeable compounds that exhibit activity in the ERF1 Redistribution assay and selectively inhibit proliferation of MAPK pathway-dependent malignant melanoma cells at similar potencies (IC(50)=< 5 microM). These compounds have drug-like structures and are negative in RAF, MEK, and ERK in vitro kinase assays. Drugs belonging to these compound classes may prove useful for treating cancers caused by excessive MAPK pathway signaling. The results also show that cell-based, high-content Redistribution screens can detect compounds with different modes of action and reveal novel targets in a pathway known to be disease relevant.     

Epidemic dynamics and antigenic evolution in a single season of influenza A

We use a mathematical model to study the evolution of influenza A during the epidemic dynamics of a single season. Classifying strains by their distance from the epidemic-originating strain, we show that neutral mutation yields a constant rate of antigenic evolution, even in the presence of epidemic dynamics. We introduce host immunity and viral immune escape to construct a non-neutral model. Our population dynamics can then be framed naturally in the context of population genetics, and we show that departure from neutrality is governed by the covariance between a strain's fitness and its distance from the original epidemic strain. We quantify the amount of antigenic evolution that takes place in excess of what is expected under neutrality and find that this excess amount is largest under strong host immunity and long epidemics.     

Characterization of alanine and malate dehydrogenases from a marine psychrophile strain PA-43

Alanine dehydrogenase (AlaDH: EC 1.4.1.1), malate dehydrogenase (MDH: EC 1.1.1.37), and glutamate dehydrogenase (EC 1.4.1.2), all NAD+ dependent, were detected in extracts from a psychrophilic bacterium, strain PA-43, isolated from a sea urchin off the Icelandic coast. Characterization tests suggested that the strain had a close relationship to Vibrio, but sequencing of part of the 16S rDNA gene placed the bacterium among Shewanella species in a constructed phylogenetic tree. The bacterium had an optimum growth temperature of 16.5 degrees C, and maximum dehydrogenase expression was obtained in a rich medium supplemented with NaCl. Both AlaDH and MDH were purified to homogeneity. AlaDH is a hexamer, with an approximate relative molecular mass of 260,000, whereas MDH is dimeric, with an apparent relative molecular mass of approximately 70,000. Both enzymes were thermolabile, and the optimum temperatures for activity were shifted toward lower temperatures than those found in the same enzymes from mesophiles, 37 degrees C for MDH and approximately 47 degrees C for AlaDH. The pH optima for AlaDH in the forward and reverse reactions were 10.5 and 9, respectively, whereas those for MDH were 10-10.2 and 8.8, respectively. Partial amino acid sequences, comprising approximately 30% of the total sequences from each enzyme, were determined for N-terminal, tryptic, and chymotryptic fragments of the enzymes. The AlaDH showed the highest similarity to AlaDHs from the psychrotroph Shewanella Ac10 and the mesophile Vibrio proteolyticus, whereas MDH was most similar to the MDHs from the mesophiles Escherichia coli and Haemophilus influenzae, with lower identity to the psychrophilic malate dehydrogenases from Vibrio 5710 and Photobacterium SS9.