Kaolinite Synthesis at 25{degrees}C

The addition of quercetin, an organic flavone (C(15)H(10)O(7)), to aqueous solutions containing silica and aluminum and adjusted to final pH's from 6.5 to 8.5 produced a 1 : 1 alumninosilicate precipitate which, after 6 to 16 months of aging in solution at 25 degrees C, contained as much as 5 percent well-formed kaolinite plates. Similar solutions containing no organic material produced relatively amorphous precipitates with the same composition and stability (standard free energy of formation = -897+/-1 kilocalories per mole) but with substantially smaller amounts of crystaline material even after 2 years of aging.     

In vivo adherence of Flavobacterium psychrophilum to mucosal external surfaces of rainbow trout (Oncorhynchus mykiss) fry

The adherence of Flavobacterium psychrophilum to surfaces of epithelial tissues has been inconclusively suggested as a mechanism, which enables the bacterium to invade the host. Hence, the present study aimed to examine the adherence of the cells of two colony phenotypes, smooth and rough, of F. psychrophilum to mucosal tissues of rainbow trout fry and to test the skin mucus as a nutrient for the growth of F. psychrophilum. Fish were immersed in water containing 10⁶ CFU mL^?1 F. psychrophilum for each colony phenotype. Mucosal tissue samples from fins, gills, skin and eyes, and swab samples from spleen and kidney were taken and inoculated onto TYES agar plates. Colony phenotypes of F. psychrophilum were identified and number of colonies counted. The results showed that cells of both phenotypes initially (0 h) adhered to all mucosal surfaces, but only the rough cells were still present on tissues 1 h post‐immersion. Both phenotypes showed a tissue tropism with the fin tissue being the most adhered. Furthermore, skin mucus promoted the growth of both colony phenotypes. We suggest that the growth of F. psychrophilum cells in skin mucus apparently facilitates the bacterial adherence to mucosal surfaces, and the subsequent invasion into the host.     

Immunohistochemical detection of SWC3, CD2, CD3, CD4 and CD8 antigens in paraformaldehyde fixed and paraffin embedded porcine lymphoid tissue

Identification of the different cell types of the immune system is important for in situ studies on the pathogenesis of infectious diseases in various animals, including the pig. Unfortunately, many monoclonal anti-leukocyte antibodies are only useful for staining frozen tissue sections with inherent poor tissue morphology, and are not readily adapted to formaldehyde fixed and paraffin embedded tissue with well preserved morphology. Seven well characterised monoclonal antibodies against porcine leukocyte antigens were tested on neutral buffered paraformaldehyde fixed and paraffin embedded porcine tissue sections using the highly sensitive tyramide signal amplification system. Combining this method with different antigen retrieval techniques enabled us to detect CD2, CD3, CD4, CD8 and SWC3 antigen expressing cells in porcine lymphoid tissue. Thus, we describe herein methods for the detection of several major cell types of the porcine immune system in fixed tissue with optimal preservation of histological details.     

Importance of soil mineral N in early spring and subsequent net N mineralisation for winter wheat following winter oilseed rape and peas in a milder climate

Abstract

Nine biennial field experiments, 2000–2004, in south Sweden, 55–56°N, with winter wheat following winter oilseed rape, peas, and oats, were used to estimate the impact of a future milder climate on winter wheat production in central Sweden, 58–60°N. The trials included studies 1) on losses during winter of soil mineral nitrogen (N_min, 0–90 cm soil), accumulated after the preceding crops in late autumn, 2) on soil N mineralisation (N_net) during the growing season of the wheat (early spring to ripeness) and 3) on grain yield and optimum N fertilisation (Opt-N rate) of the wheat. Average N_min in late autumn following winter oilseed rape, peas, and oats was 68, 64, and 45 kg ha^?1, respectively, but decreased until early spring. Increased future losses of N_min during the winter in central Sweden due to no or very short periods with soil frost should enhance the demand for fertiliser N and reduce the better residual N effect of winter oilseed rape and peas, compared with oats. Their better N effect will then mainly depend on larger N_net (from March to maturity during the winter wheat year). Owing to more plant-available soil N (mainly as N_net) Opt-N rates were lower after oilseed rape and peas than after oats despite increased wheat yields (700 kg ha^?1) at optimum N fertilisation. In addition to these break crop effects, a milder climate should increase winter wheat yields in central Sweden by 2000–3000 kg ha^?1 and require about 30–45 kg ha^?1 more fertiliser N at optimum N fertilisation than the present yield levels. Increased losses and higher N fertilisation to the subsequent winter wheat in future indicates a need for an estimation of the residual N effect at the individual sites, rather than using mean values as at present, to increase N efficiency.     

Direct experimental evidence for the contribution of lime to CO2 release from managed peat soil

Liming is a common management practice used to achieve optimum pH for plant growth in agricultural soils. Addition of lime to the soil, however, may cause CO₂ release when the carbonates in lime dissolve in water. Although lime may thereby constitute a significant carbon source, especially under acidic soil conditions, experimental data on the CO₂ release are lacking so far. We conducted a split-plot experiment within a cut-away peatland cultivated with a bioenergy crop (reed canary grass, Phalaris arundinacea L.) with lime and fertilizer treatments to determine effects of lime on the CO₂ emissions from soil and to better understand mechanisms underlying liming effects. Carbon dioxide release was measured over two growing seasons in the field after liming, and complementary laboratory studies were conducted. To differentiate CO₂ derived from lime and biotic respiration the δ¹³C of CO₂ released was determined and the two-pool mixing model was applied. The results showed that lime may contribute significantly to CO₂ release from the soil. In the laboratory, more than 50% of CO₂ release was attributable to lime-carbonates during short-term incubation. Lime-derived CO₂ emissions were much lower in the field, and were only detected during the first (2–4) months after the application. However, a maximum of 12% of monthly CO₂ emissions from the cultivated peatland originated from the lime. Biotic respiration rates were similar in limed and unlimed soils, suggesting that higher pH did not, at least in the short-term, increase carbon losses from cultivated peat soils. Additional fertilization and acidification did not contribute to further CO₂ release from the lime. According to our first estimations about one sixth of the lime applied would be released as CO₂ from the managed peatland, with all lime-derived emissions occurring during the first year of application (equivalent to about 4.6% of the total annual CO₂ losses from the soil in the first year). This suggests that the mass-balance approach as proposed by the IPCC Tier 1 methodology, which assumes that all carbon in lime ends up as CO₂ in the atmosphere, overestimates the emissions from lime. Our study further shows that there is a great risk to overestimate heterotrophic microbial activity in limed soils by measuring the CO₂ release without separating abiotic and biotic CO₂ production.     

Mutational robustness of ribosomal protein genes

The distribution of fitness effects (DFE) of mutations is of fundamental importance for understanding evolutionary dynamics and complex diseases and for conserving threatened species. DFEs estimated from DNA sequences have rarely been subject to direct experimental tests. We used a bacterial system in which the fitness effects of a large number of defined single mutations in two ribosomal proteins were measured with high sensitivity. The obtained DFE appears to be unimodal, where most mutations (120 out of 126) are weakly deleterious and the remaining ones are potentially neutral. The DFEs for synonymous and nonsynonymous substitutions are similar, suggesting that in some genes, strong fitness constraints are present at the level of the messenger RNA.     

Technical note: time-resolved immunofluorometric assay for growth hormone in ruminants

A noncompetitive, time-resolved immunofluorometric assay (TRIFMA) was developed using a selected pair of monoclonal antibodies (mab) raised against recombinant bovine GH, with the catching mab immobilized on microtiter plate wells and the detection mab labeled with Eu3+ as a tracer, arranged as a sandwich. Plates were coated with mab1.15 (680 ng/well) using a phosphate buffer (pH 4.9), and then blocked with assay buffer containing 1% (wt/vol) BSA. The assay procedure involved incubation of 50 microL of sample (plasma or serum) and 200 microL of assay buffer containing 25 ng of mab1.2-Eu3+ conjugate for 4 h at 25 degrees C. Plates were then washed six times, incubated for 5 to 10 min with 250 microL of enhancement solution, and fluorescence read with a time-resolved fluorometer. The sensitivity of the assay was 0.1 ng/mL, and the working range was 0.2 to 200 ng/mL. Recovery of quantitative amounts of bovine GH added to plasma samples was close to 100%. Cross-reactivity with other bovine pituitary hormones or with GH from nonbovidae or cervidae species was not significant. Intra- and interassay CV during routine operation was 4.4 and 10.7%, respectively (mean = 3.54 ng/mL). Plasma concentrations of bovine GH determined by TRIFMA correlated closely (r2 > or = 0.93) with RIA results, with a conversion ratio of 0.62 when the higher specificity of the monoclonal antibodies was taken into account. The TRIFMA is a reliable alternative to RIA methods because the assay employs no radiolabeled or hazardous chemicals, delivers results rapidly, and has little risk of down periods.