Innovations in the diagnosis of infectious agents of veterinary importance--a seven-league boots fairy tale?

The development of infectiological diagnostic methodology is characterized by saltatoric, revolutionary leaps. Improved sensitivity and specificity, reduction of analytical sample volumes, increased sample throughput, reduced test complexity associated with improved user friendliness are hallmarks of such developments. Apart from purely technical amenities some innovations also enable conceptual adaptations of monitoring strategies. These include the use of diagnostic assays on-site, e. g. in the stable or in the practice. Selecting the assay which is most "fit-for-purpose" among the plethora of sometimes loudly advertised new technology may be difficult even for the well trained and experienced diagnostician. Standardized licensing procedures for infectiological diagnostic kits may be required to guarantee minimal norms of diagnostic quality. At the same time availability of a diversity of diagnostic kits is needed to broaden the base of diagnostic reliability. New infectiological diagnostic innovations will only be useful if the (expanded) results are transposable to the improved practical care of companion animals and farm animal holdings. In addition, the judgment of results, e. g. for epidemiological plausibility, remains a very important issue even when the test results have been produced by novel high technology diagnostics.     

Straw management, crop rotation, and nitrogen source effect on wheat grain yield and nitrogen use efficiency

Nitrogen fertilizer management from different sources and annual crop rotations are important components of wheat (Triticum aestivum L.) production systems, especially where air and soil quality issues have prompted a search for alternatives to wheat straw burning. This study examined the effects of two different wheat straw management options (burning and incorporation by tillage), three crop rotations [wheat-sesbania (Sesbania spp.), wheat–maize (Zea mays L.), and wheat-clean fallow] and three N sources (urea, chicken manure, and urea plus chicken manure) on wheat grain yield and N use efficiency. The experiment was conducted as split–split plot treatment arrangement with three replications for eight wheat cropping seasons in the state of Sonora, Mexico. Results indicated that both wheat grain yield and N use efficiency were higher with burning than incorporation of wheat straw and with fertilization with urea or urea plus chicken manure than chicken manure alone. As shown by the crop rotation-by-straw management interaction, planting sesbania following incorporation of the straw by tillage produced comparable grain yields to straw burning treatment. In contrast, wheat in annual rotation with maize produced the lowest wheat grain yield and N use efficiency irrespective of the wheat straw management and N source applied.     

Identification of fatty acid esters of pectenotoxin-2 seco acid in blue mussels (Mytilus edulis) from Ireland

Pectenotoxins from marine dinoflagellates of the genus Dinophysis are rapidly hydrolyzed by many shellfish to give pectenotoxin-2 seco acid, which isomerizes to 7-epi-pectenotoxin-2 seco acid. Three series of fatty acid esters of pectenotoxin-2 seco acid (PTX-2 seco acid) and 7-epi-PTX-2 seco acid were detected by LC-MS analysis of extracts from blue mussels (Mytilus edulis) from Ireland. The locations of the fatty acid ester linkages were identified by a combination of LC-MSn in positive- and negative-ion modes, LC-MS analysis of the products from reaction of the esters with sodium periodate, and NMR analysis of purified samples of the two most abundant ester derivatives. The 37-O-acyl esters of PTX-2 seco acid were the most abundant, followed by the corresponding 11-O-acyl esters, accompanied by low levels of the 33-O-acyl esters. The most abundant fatty acid esters in the fractionated sample were, in order, the 16:0, 22:6, 14:0, 16:1, 18:4, and 20:5 fatty acids, although a wide array of other PTX-2 seco acid fatty acid esters were also present at low levels.     

Effects of an arbuscular mycorrhizal fungus and two plant-parasitic nematodes on<Emphasis Type="Italic"> Musa</Emphasis> genotypes differing in root morphology

In this study, the effect of an arbuscular mycorrhizal fungus (AMF) and two migratory endoparasitic nematodes on Musa plant growth, including the root system, were examined. In addition, the AMF-nematode interaction was studied. Seven Musa genotypes with different root systems were selected. Based on their relative mycorrhizal dependency, two genotypes (Calcutta 4 and Obino l'Ewai) were selected for AMF-nematode interaction studies. The experiments were performed under greenhouse conditions. Mycorrhization with Glomus mosseae resulted in a significantly better plant growth even in the presence of nematodes. The effect of AMF on the root system was genotype-dependent and seemed to be related to the relative mycorrhizal dependency of the genotype. The nematodes also affected the root system, decreasing branching. Nematode population densities were significantly reduced in the presence of AMF, except for Pratylenchus coffeae in Obino l'Ewai. In the root system, it appeared that the decreased branching caused by the nematodes was counterbalanced by the increased branching caused by the AMF.     

Effect of High‐Molecular‐Weight Glutenin Subunit Allelic Composition on Wheat Flour Tortilla Quality

Wheat cultivars possessing quality attributes needed to produce optimum quality tortillas have not been identified. This study investigated the effect of variations in high‐molecular‐weight glutenin subunits encoded at the Glu‐1 loci (Glu‐A1, Glu‐B1, and Glu‐D1) on dough properties and tortilla quality. Flour protein profiles, dough texture, and tortilla physical quality attributes were evaluated. Deletion at Glu‐D1 resulted in reduced insoluble polymeric protein content of flour, reduced dough compression force, and large dough extensibility. These properties produced very large tortillas (181 mm diameter) compared with a control made with commercial tortilla wheat flour (161 mm). Presence of a 7 + 9 allelic pair at Glu‐B1 increased dough strength (largest compression force, reduced extensibility, and small‐diameter tortillas). Deletion at Glu‐A1 produced large tortillas (173 mm) but with unacceptable flexibility during storage (score <3.0 at day 16). In general, presence of 2* at Glu‐A1, in combination with 5 + 10 at Glu‐D1, produced small‐diameter tortillas that required large force to rupture (tough texture). Presence of 2 + 12 alleles instead of 5 + 10 at Glu‐D1 produced tortillas with a good compromise between diameter (>165 mm) and flexibility during storage (>3.0 at day 16). These allele combinations, along with deletion at Glu‐D1, show promise for tortilla wheat development.     

Bioassay-directed Fractionation of Organic Extracts of Marine Surface Sediments from the North and Baltic Sea - Part II: Results of the biotest battery and development of a biotest index (8 pp)

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Part I: Determination and identification of organic pollutants Part II: Results of the biotest battery and development of a biotest index

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Preamble. This series of two papers presents the results of an interdisciplinary research project (ISIS) dealing with bioassay-directed fractionation of marine sediment extracts. Part I presents the extraction and fractionation procedure as well as the results of chemical analysis, including non-target analysis of sediments. Part II describes the results of the biotest battery in relation to chemicals possibly causing parts of the observed effects. A biotest index is used to compare the toxicities of the samples.

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AUTHORS / AFFILIATIONS Ninja Reineke (3), Werner Wosniok (4), Dirk Danischewski (1), Heinrich Hühnerfuss (3), Angelika Kinder (5), Arne Sierts-Herrmann (5), Norbert Theobald (2), Hans-Heinrich Vahl (6), Michael Vobach (1), Johannes Westendorf (6) and Hans Steinhart (5).

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(1) Federal Research Centre for Fisheries, Institute for Fishery Ecology, Palmaille 9, 22767 Hamburg, Germany (2) Federal Maritime and Hydrographic Agency, Bernhard-Nochtstr. 78, 20359 Hamburg, Germany (3) University of Hamburg, Institute for Organic Chemistry, Martin-Luther-King-Platz 6, 20146 Hamburg, Germany (4) University of Bremen, Institute of Statistics, Bibliothekstr. 1, 28334 Bremen, Germany (5) University of Hamburg, Institute for Food Chemistry, Grindelallee 117, 20146 Hamburg, Germany (6) University of Hamburg, University Hospital Hamburg-Eppendorf, Department for Toxicology, Vogt-Kölln-Str. 30, 22527 Hamburg, Germany (7) Eurofins Wiertz-Eggert-Jörissen, Stenzelring 14b, 21107 Hamburg, Germany

Goal, Scope and Background

The ecological relevance of contaminants in mixtures is difficult to assess, because of possible interactions and due to lacking toxicity data for many substances present in environmental samples. Marine sediment extracts, which contain a mixture of environmental contaminants in low concentrations, were the object of this study. The extracts were investigated with a set of different biotests in order to identify the compound or the substance class responsible for the toxicity. For this goal, a combination of biotests, biotest-directed fractionation and chemical analysis has been applied. Further on, a strategy for the development of a biotest index to describe the toxicity of the fractions without a prior ranking of the test results is proposed. This article (Part II) focuses on the biological results of the approach.

Methods

The toxicological potential of organic extracts of sediments from the North Sea and the Baltic Sea was analyzed in a bioassay-directed fractionation procedure with a set of biotests: luciferase reporter gene assays on hormone receptor and Ah receptor, arabinose resistance test, fish embryo test (Danio rerio), comet assay, acetylcholinesterase inhibition test, heat-shock protein 70 induction, oxidative stress and luminescence inhibition test (Vibrio fischeri). The test results provided the basis for the calculation of a biotest index by factor analysis to compare the toxicity of the samples and fractions.

Results and Discussion

Results of 11 biotests on different fractionation levels of the samples were described and discussed with regard to the occurrence of contaminants and their toxic potentials. Polychlorinated biphenyls, polycyclic aromatic hydrocarbons, quinones, brominated indoles and brominated phenols were in the focus of interest. A biotest index was constructed to compare the toxic responses in the samples and to group the biotest results.

Conclusion

The procedure presented in this study is well suited for bioassay-directed fractionation of marine sediment extracts. However, in relatively low contaminated samples, high enrichment factors and sufficient fractionation is necessary to allow identification of low concentrations of contaminants which is required to link effects and possible causes. In the present case, the relation between substances and effects was difficult to uncover due to relatively low concentrations of pollutants compared to the biogenic matrix and to the remaining complexity of the fractions. The results, with respect to the brominated phenols and indoles in the samples, highlight the successful use of bioassay directed fractionation in the case of high concentrations and high toxicity.

Recommendation and Outlook

In general, it has been shown that a marine risk assessment requires focusing on the input of diffuse sources and taking into account the fact of mixture toxicity. Effects resulting from biogenic substances will make the assessment of the influence of anthropogenic substances even more difficult.     

Model calculations of water dynamics in lysimeters filled with granular industrial wastes

A three‐year long lysimeter experiment with a fine‐grained aluminum (Al) recycling by‐product and a mixture of this by‐product and a coal combustion waste was conducted. The wastes were proposed as possible soil substitutes in an engineered surface barrier covering a potash mining residue mount. To evaluate the suitability of the wastes as surface barrier material, their hydrological behavior under field conditions must be known. Lysimeter experiments provide one means to study the hydrological behavior of soils or soil‐like materials. However, it is difficult to estimate the long‐term hydrological behavior from short‐term lysimeter studies. The present study was conducted therefore to derive from short‐term lysimeter observations the long‐term hydrological behavior of the two waste materials. The lysimeter data were used to calibrate the one‐dimensional soil water flow model HYDRUS‐1D. With the calibrated model, hydrological simulations for the site of the residue mount were carried out for a period of 31 yr. Calculated long‐term annual seepage from the lysimeters was 237 mm for the pure Al waste and 186 mm for the mixture, or 39% and 24% of the average annual precipitation (764 mm). The average discharge of the bare mount is 482 mm or 63%. We conclude that a soil cover could considerably reduce the discharge and that the mixture is better suited as surface barrier than the pure Al waste.     

Nuclear magnetic resonance spectroscopy and chemometrics to identify pine nuts that cause taste disturbance

Nontargeted 400 MHz (13)C and (1)H nuclear magnetic resonance (NMR) spectroscopy was used in the context of food surveillance to reveal Pinus species whose nuts cause taste disturbance following their consumption, the so-called pine nut syndrome (PNS). Using principal component analysis, three groups of pine nuts were distinguished. PNS-causing products were found in only one of the groups, which however also included some normal products. Sensory analysis was still required to confirm PNS, but NMR allowed the sorting of 53% of 57 samples, which belong to the two groups not containing PNS species. Furthermore, soft independent modeling of class analogy was able to classify the samples between the three groups. NMR spectroscopy was judged as suitable for the screening of pine nuts for PNS. This process may be advantageous as a means of importation control that will allow the identification of samples suitable for direct clearance and those that require further sensory analysis.     

Soil nutrient supply and biomass production in a mixed forest on a skeleton‐rich soil and an adjacent beech forest

In the natural forest communities of Central Europe, beech (Fagus sylvatica L.) predominates in the tree layer over a wide range of soil conditions. An exception with respect to the dominance of beech are skeleton‐rich soils such as screes where up to 10 broad‐leaved trees co‐exist. In such a Tilia‐Fagus‐Fraxinus‐Acer‐Ulmus forest and an adjacent mono‐specific beech forest we compared (1) soil nutrient pools and net nitrogen mineralization rates, (2) leaf nutrient levels, and (3) leaf litter production and stem increment rates in order to evaluate the relationship between soil conditions and tree species composition. In the mixed forest only a small quantity of fine earth was present (35 g l^—1) which was distributed in patches between basalt stones; whereas a significantly higher (P < 0.05) soil quantity (182 g l^—1) was found in the beech forest. In the soil patches of the mixed forest C and N concentrations and also concentrations of exchangeable nutrients (K, Ca, Mg) were significantly higher than in the beech forest. Net N mineralization rates on soil dry weight basis in the mixed forest exceeded those in the beech forest by a factor of 2.6. Due to differences in fine earth and stone contents, the volume related soil K pool and the N mineralization rate were lower in the mixed forest (52 kg N ha^—1 yr^—1, 0—10 cm depth) than in the beech forest (105 kg N ha^—1 yr^—1). The leaf N and K concentrations of the beech trees did not differ significantly between the stands, which suggests that plant nutrition was not impaired. In the mixed forest leaf litter fall (11 %) and the increment rate of stem basal area (52 %) were lower than in the beech forest. Thus, compared with the adjacent beech forest, the mixed forest stand was characterized by a low volume of patchy distributed nutrient‐rich soil, a lower volume related K pool and N mineralization rate, and low rates of stem increment. Together with other factors such as water availability these patterns may contribute to an explanation of the diverse tree species composition on Central European screes.     

Producing native seed of Stipa lagascae R. and Sch. inPresaharian Tunisia: high yields, high genetic variation orboth?

Genetic shifts are predicted to be greatest when highly diversematerial originating from an arid environment is bulk multiplied inirrigated and fertilised conditions. This is a key problem if largequantities of quality native seed are needed for restorationpurposes. In Presaharian Tunisia, we investigated two intermingledyield aspects of irrigated seed multiplication of Stipalagascae R. and Sch.: (1) nitrogen fertilisationand (2) selection for vegetative vigour. We set up twocontiguous nitrogen fertilisation trials, one withnon-selected seeds (from a nearby locality), and onewith selected seeds (48 clonal lines seeded in replicatedrows), and compared straw yields. Both selected and unselectedmaterial did not respond to more than 50 kg of nitrogenper ha. Unfertilised yields of both trials were similar,but the straw yield of fertilised selected material was more than100% higher than the straw yield of fertilised unselectedmaterial. In the trial with selected material, we collected straw andleaf yields per row, and scored each row for vigour and for phenologyduring three successive growing seasons. Genotypic differences inseed output and in mortality were large and mainly driven byvariation in potential straw for leaf ratios.The compromise between high yields and high genetic variation couldconsist of the following principles. (1) Create driergrowing conditions so that the actual straw forleaf ratio stays below the genotypic potential. (2) Useinstead fertilisation and selection to enhance seed yields.(3) Avoid bulk multiplication.