Susceptibility of Escherichia coli and Enterococcus faecium isolated from pigs and broiler chickens to tetracycline degradation products and distribution of tetracycline resistance determinants in E. coli from food animals

One hundred Escherichia coli isolates from diseased and healthy pigs, cattle and broiler chickens were screened for the presence of tetracycline resistance genes tet(A), (B), (C), (D) or (E). The tet(A) gene was the most abundant (71% of the 100 isolates) followed by tet(B) (25%). The predominance of tet(A) and tet(B) applied to all three animal species, and there was no difference between the distribution of tet(A) and tet(B) genes among non-pathogenic and pathogenic E. coli in any of the animal species. The susceptibility of 20 of these isolates together with 10 tetracycline sensitive E. coli and 18 tetracycline resistant and 10 sensitive Enterococcus faecium to tetracyclines and tetracycline degradation products was determined. The resistant isolates showed reduced resistance to anhydrotetracycline, 4-epi-anhydrotetracycline, anhydrochlortetracycline and 4-epi-anhydrochlortetracycline. In general both the tetracycline resistant and susceptible E. faecium were more susceptible to the compounds tested than E. coli.     

Effects of long-term contrasting lime and phosphorus applications on barley grain yield,root growth and abundance of mycorrhiza

Lime and phosphorus (P) applications are common agricultural management practices. Our aim was to quantify the effects of long-term application practices on root growth and abundance of arbuscular mycorrhizal fungi (AMF) under field conditions. We assessed the effects of lime and P fertilizer applications on barley yield, root growth and AMF abundance in 2016. Treatments were no, low, medium and high liming rate corresponding to application of 0, 4, 8 and 12 Mg lime ha⁻¹ every 5–9 years since 1942 combined with no or yearly application of 15.6 kg P ha⁻¹ since 1944. At harvest, grain yield, root intensity (core-break) and AMF abundance at different soil depths were estimated. Root development was monitored during early growth with minirhizotrons in treatments receiving low, medium and high liming rates and P fertilization. A quadratic model relating grain yield to liming rate estimated yields to peak at 6.4 Mg lime ha⁻¹ with yields of 4.2 and 3.2 Mg grain ha⁻¹ with and without P fertilization, respectively. Low and medium liming rates resulted in greater AMF abundance, especially in the no P treatments. During early growth in P-fertilized treatments, 77% and 65% more roots developed in the soil profile when treated with medium and high liming rate, respectively, compared to low liming rate. We conclude that long-term application of lime in soils receiving yearly P fertilization improved conditions for root growth in soil layers below 30 cm, but at the high liming rate, this did not translate into higher yield.     

Fertilizer ammonium : nitrate ratios determine phosphorus uptake by young maize plants

We investigated the interacting effects of inorganic nitrogen and the main inorganic phosphorus form in dairy manure (dicalcium phosphate, CaHPO₄) on growth, nutrient uptake, and rhizosphere pH of young maize plants. In a pot experiment, three levels of CaHPO₄ (0, 167, and 500 mg P pot^?1) were combined with nitrogen (637 mg N pot^?1) applied at five NH₄‐N : NO₃‐N ratios (0 : 100, 25 : 75, 50 : 50, 75 : 25, and 100 : 0) and a nitrification inhibitor in a concentrated layer of a typical acid sandy soil from Denmark. ¹⁵N‐labeled NH₄‐N was applied to differentiate the role of nitrification and to partition nitrogen uptake derived from NH₄‐N. Among treatments including nitrogen, shoot biomass, rooting and phosphorus uptake were significantly higher at the five‐leaf stage when CaHPO₄ was applied with NH₄‐N : NO₃‐N ratios of 50 : 50 and 75 : 25. In these treatments, rhizosphere pH dropped significantly in direct proportion with NH₄‐N uptake. The fertilizers in the concentrated layer had a root‐inhibiting effect in treatments without phosphorus supply and in treatments with pure NO₃‐N or NH₄‐N supply. Increased nitrogen uptake as NH₄‐N instead of NO₃‐N reduced rhizosphere pH and enhanced acquisition of applied CaHPO₄ by young maize plants, which may have positive implications for the enhanced utilization of manure phosphorus.     

Phenolic Acids of the Two Major Blueberry Species in the US Market and Their Antioxidant and Anti-inflammatory Activities

Plant Foods for Human Nutrition - Highbush (cultivated) and lowbush (wild) are the two major blueberry species in the US market. Eight phenolic acids were detected and quantified from these two...     

Long‐term effects of animal manure and mineral fertilizers on phosphorus availability and silage maize growth

A better appraisal of the plant availability of soil phosphorus (P) added with animal manure is crucial to alleviate environmental impacts from over‐application of P. This study compares the availability of P to maize in the Askov long‐term experiments using unmanured plots and plots receiving corresponding rates of nitrogen (N), P and potassium (K) in mineral fertilizers or manure. Total‐P and water extractable P (Pw) in soil, and plant height, dry weight, P concentration and P uptake were determined in early August. Final yields were determined in late October. Soil Pw was similar for plots receiving corresponding rates of P in mineral fertilizer or manure form. With a strong relationship between Pw, and maize growth and final yields, Pw was a reliable indicator of P availability to maize. Plant dry weight, P concentration and P uptake in early August were 23%, 8% and 31% higher, respectively, for maize grown on soil receiving manure compared with mineral fertilizer, while final maize yield in late October was 13% higher. Plant height and dry weight determined in early August suggested that maize development at this growth stage defined final maize yield. We conclude that the availability of P was similar after long‐term application of corresponding rates of P in animal manure and mineral fertilizers, and that animal manure improves the growth of maize compared to mineral fertilizers. This is ascribed to micronutrients and residual N effects from previous additions of manure.     

A new method for determining the metabolic activity of specific bacterial populations in soil using tritiated leucine and immunomagnetic separation

 A new assay, using immunomagnetic separation and uptake of tritiated leucine ([³H]-Leu), was developed for measuring the in situ metabolic activity of specific bacterial populations in soil. Such assays are needed to assess the role individual species play in diverse microbial soil communities. The method was optimized using Pseudomonas putida KT2440 : :Tc⁺/TOL::gfp inoculated into soil microcosms. Inoculated soil samples were incubated with [³H]-Leu followed by an immunomagnetic separation to recover the target bacteria. Radiolabel incorporated by the target bacteria was then measured. Incubation time with immunomagnetic beads was not critical for optimal target cell recovery, but samples needed to be washed at least 5 times during the immunomagnetic separation to reduce unspecific binding of the indigenous soil bacteria to the magnetic beads. Soil absorption of the polyclonal antibody further reduced this unspecific binding, resulting in <1% contamination by indigenous soil bacteria relative to numbers of recovered target cells. The assay was tested by investigating the effect of different incubation temperatures on the metabolic activity of the target cells. As expected, a linear relationship between activity and temperature was observed, demonstrating the sensitivity of the assay. The method was applied to compare activities of the target strain in bulk soil and in the rhizosphere of barley. Contrary to what was anticipated, no significant difference in metabolic activity was observed. Received: 4 November 1999     

Quantification of anthocyanins in commercial black currant juices by simple high-performance liquid chromatography. Investigation of their pH stability and antioxidative potency

Quantitative determinations of the four black currant anthocyanins, cyanidin 3-O-beta-glucoside, cyanidin 3-O-beta-rutinoside, delphinidin 3-O-beta-glucoside, and delphinidin 3-O-beta-rutinoside, were achieved in black currant juices by a rapid and sensitive high-performance liquid chromatographic (HPLC) method. The method was validated, and quantification of anthocyanins in 13 commercially available black currant beverages was demonstrated. To optimize the handling of anthocyanin-containing samples, the pH-dependent stability of the anthocyanins was investigated. Four anthocyanins were incubated for 24 h in aqueous solutions at 13 different pH levels between 0.6 and 5.2, after which the samples were analyzed by HPLC. More than 90% of each anthocyanin remained intact up to pH 3.3. At pH 3.8 a local minimum in stability was detected, and at pH >4.5 the stability rapidly decreased. The antioxidant capacity of all 13 black currant juices was investigated by TEAC and FRAP, and the antioxidant potential of both the anthocyanin and the vitamin C contents in the juices was evaluated. This indicated that <70% of the antioxidant capacity of the juices could be attributed to the anthocyanin content or to vitamin C, signifying that other very potent antioxidants are present in commercial black currant juices.     

An enzyme-linked immunosorbent assay for detection of avian influenza virus subtypes H5 and H7 antibodies

Background

Avian influenza virus (AIV) subtypes H5 and H7 attracts particular attention because of the risk of their potential pathogenicity in poultry. The haemagglutination inhibition (HI) test is widely used as subtype specific test for serological diagnostics despite the laborious nature of this method. However, enzyme-linked immunosorbent assays (ELISAs) are being explored as an alternative test method.H5 and H7 specific monoclonal antibodies were experimentally raised and used in the development of inhibition ELISAs for detection of serological response specifically directed against AIV subtypes H5 and H7. The ELISAs were evaluated with polyclonal chicken anti-AIV antibodies against AIV subtypes: H1N2, H5N2, H5N7, H7N1, H7N7, H9N9, H10N4 and H16N3.

Results

Both the H5 and H7 ELISA proved to have a high sensitivity and specificity and the ELISAs detected H5 and H7 antibodies earlier during experimental infection than the HI test did. The reproducibility of the ELISA’s performed at different times was high with Pearson correlation coefficients of 0.96-0.98.

Conclusions

The ELISAs are a potential alternative to the HI test for screening of large amounts of avian sera, although only experimental sera were tested in this study.     

Availability of potassium in biomass combustion ashes and gasification biochars after application to soils with variable pH and clay content

The expansion of the bioenergy sector and adoption of novel thermal conversion technologies produce increasingly large amounts of biomass ashes and biochars. Before returning such products to agricultural soil, the plant availability of nutrients when mixing with soil should be assessed. The objective of this study was to evaluate the potassium (K) availability in various types of biomass ashes and gasification biochars (GBs) derived from straw, wood, sewage sludge and poultry manure when mixed with soil. A 16-week incubation study was conducted with three contrasting soils of variable pH (5.8–7.8) and clay contents (3–17%). Exchangeable K in the product-soil mixture was determined, and the K recovery rate from the applied products varied from 31 to 86%. The relative recovery compared to applied KCl was used to indicate K availability and was 50–86% across all soils, but lower for two sewage sludge-based GBs in the alkaline soil. Most of the K release occurred within the first week, with insignificant release thereafter. Wet storage of GB did not affect K availability. We conclude that the tested biochars and ashes can be used as K fertilisers with slightly lower short-term availability than KCl.     

Comparison of chemical and cell-based antioxidant methods for evaluation of foods and natural products: generating multifaceted data by parallel testing using erythrocytes and polymorphonuclear cells

The objective of this study was to compare three tests frequently used for evaluation of antioxidant potential in natural products: (1) oxygen radical absorbance assay (ORAC), (2) cell-based antioxidant protection in an erythrocyte model (CAP-e), and (3) reactive oxygen species formation in polymorphonuclear cells (ROS PMN). The methods were applied to four natural products, all containing antioxidants capable of entering and protecting cells in the CAP-e assay. The magnitude of this effect was not directly correlated to the ORAC value of each product. Furthermore, the products showed different effects in the ROS PMN assay. Acai provided strong inhibition of ROS formation, indicating anti-inflammatory properties. In contrast, Immunel and EpiCor mildly enhanced ROS formation, suggesting activation of the innate immune response. HA Joint Formula showed a complex, nonlinear dose-response in the ROS PMN assay. This illustrates that complex natural products may have similar antioxidant properties but different effects on human cells. Cell-based antioxidant protection is addressed best in the CAP-e assay, since some natural products contain compounds that may provoke cellular signaling in other cell types. The PMN cell type is a useful model for assessment of overall anti-inflammatory versus immune supportive properties of a product. The sequential use of the three methods serves to bridge analytical and biological testing methods.