Simple and Rapid Quality Control of Sulfated Glycans by a Fluorescence Sensor Assay—Exemplarily Developed for the Sulfated Polysaccharides from Red Algae Delesseria sanguinea

Sulfated polysaccharides (SP) from algae are of great interest due to their manifold biological activities. Obstacles to commercial (especially medical) application include considerable variability and complex chemical composition making the analysis and the quality control challenging. The aim of this study was to evaluate a simple microplate assay for screening the quality of SP. It is based on the fluorescence intensity (FI) increase of the sensor molecule Polymer-H by SP and was originally developed for direct quantification of SP. Exemplarily, 65 SP batches isolated from the red alga Delesseria sanguinea (D.s.-SP) and several other algae polysaccharides were investigated. Their FI increase in the Polymer-H assay was compared with other analytical parameters. By testing just one concentration of a D.s.-SP sample, quality deviations from the reference D.s.-SP and thus both batch-to-batch variability and stability can be detected. Further, structurally distinct SP showed to differ in their concentration-dependent FI profiles. By using corresponding reference compounds, the Polymer-H assay is therefore applicable as identification assay with high negative predictability. In conclusion, the Polymer-H assay showed to represent not only a simple method for quantification, but also for characterization identification and differentiation of SP of marine origin.     

Sulfonamides Leach from Sandy Loam Soils Under Common Agricultural Practice

Sulfonamide antibiotics can enter agricultural soils by fertilisation with contaminated manure. While only rough estimations on the extent of such applications exist, this pathway results in trace level contamination of groundwater. Therefore, we studied the transport of three sulfonamides in leachates from field lysimeters after application of a sulfonamide-contaminated liquid manure. In a 3-year period, the sulfonamides were determined in 64% to 70% of all leachate samples at concentrations between 0.08 to 56.7 µg L^?1. Furthermore, sulfonamides were determined in leachates up to 23 months after application, which indicated a medium- to long-term leaching risk. Extreme dry weather conditions resulted in highest dislocated amounts of sulfonamides in two of the three treatments. Furthermore, soil management such as tillage and cropping affected the time between application and breakthrough of sulfonamides and the intra-annual distribution of sulfonamide loads in leachates. Although the total sulfonamide leaching loads were low, the concentrations exceeded the limit value of the European Commission of 0.1 µg biocide L^?1 in drinking water in more than 50% of all samples. Furthermore, the medium-term mean concentration of the sulfonamides ranged from 0.08 and 4.00 µg L^?1, which was above the limit value of the European Commission in 91 out of 158 samples. Therefore, sulfonamides applied to soils in liquid manure under common agricultural practice may cause environmental and health risks which call for a setting up of more long-term studies on the fate of antibiotics.     

Fast and efficient discrimination of the Isotoma viridis group (Insecta: Collembola) by PCR-RFLP

Identification of collembolan species is generally based on specific morphological characters, such as chaetotaxy and pigmentation pattern. However, some specimens do not match to described characters because these refer to adult specimens, often of one specific sex, or the characters are highly variable in adults (e.g. pigmentation, setae or furcal teeth). Isozymes have frequently assisted species discrimination, and also these may vary with developmental stage or environmental conditions. For identification of single species of the Isotoma viridis group, we present both direct sequencing of the cytochrome oxidase subunit II (COII) gene and a simple DNA-based molecular method.

Five PCR primers amplifying the COII region (717 bp) of the mitochondrial DNA were used. The sequences clearly separated the species I. viridis, I. riparia and I. anglicana, irrespective of colour varieties within the first species. DNA amplification products of different species can also be distinguished by digestion with restriction endonucleases, followed by gel electrophoresis for separation of fragments. This restriction fragment length polymorphism (RFLP), obtained after digestion with the endonucleases TaqI, VspI, MvaI and Bsp143I, revealed specific fragments that separated the three species from each other. Since restriction enzymes are sensitive to single base mutations, we suggest to use a combination of enzymes with at least two species-specific restriction sites when using the RFLP technique. For the I. viridis complex, VspI and Bsp143I appear to be an appropriate combination.     

Food choice and reproductive success of Folsomia candida feeding on copper-contaminated mycelium of the soil fungus Alternaria alternata

We examined collembolan food preference for fungal mycelium grown on copper-contaminated medium, and the relationship between copper content, food selectivity and collembolan fitness when fed contaminated mycelium.To clarify whether collembolan food selectivity is related to fitness parameters, Folsomia candida were fed mycelium of the dark-pigmented fungus Alternaria alternata grown on medium with different copper concentrations. Copper-contaminated food (fungus grown on 50, 125, 250 and 500 μg Cu g^?1 medium, fresh wt.) was offered together with untreated food for 4 weeks. F. candida fed selectively on the provided mycelium and discriminated clearly between mycelium grown on high and low levels of contamination, distinctly preferring fungus grown on medium with a total copper concentration of 50 and 125 μg g^?1. In contrast, fungus grown on highly contaminated medium (250 and 500 μg g^?1) was avoided. Collembolan food preference generally matched fitness parameters. Reproduction was significantly affected by the total copper concentration of the fungal growth medium. When fed their preferred mycelium, collembolan reproduction was enhanced, whereas a diet of highly contaminated mycelium (250 or 500 μg g^?1) resulted in a strong decrease in reproduction. Adult survival was affected only marginally. Even though heavy metal contamination is a potential stress factor for many soil microarthropods, F. candida is able to discriminate between high and low quality food sources, and even benefits from moderately elevated copper concentrations.     

Bottom-up down from the top: Honeydew as a carbon source for soil organisms

Honeydew produced by aphids is a well-studied food source for mutualistic ants. Although considerable amounts drop onto the soil surface there are hardly any studies dealing with the potential effects of this additional C source for soil organisms, particularly those living at the soil surface. We studied this question by simulating honeydew and investigating plots under Populus canescens formerly infested with aphids on two ruderal sites differing in soil organic matter content and vegetation cover. The simulated honeydew input increased microbial biomass with some delay in week 10. Both natural and simulated honeydew raised the activity densities of an epigeic Collembola taxon, the Bourletiellidae, but not of the dominant Hemisotoma thermophila. The honeydew effect varied over time and with the site, and was more pronounced at the nutrient-poor site for Collembola, but not for microbial biomass. Ant consumption reduced the amount of honeydew reaching the soil surface by 50%. The activity density of H. thermophila was negatively related to ants and spiders, hinting at a top-down control. Honeydew clearly acts as a bottom-up force for soil organisms, which is however only part of the complex network connecting the food webs of primary producers and decomposers.     

Comparison of computer assisted surgery with conventional technique for treatment of abaxial distal phalanx fractures in horses: an in vitro study

Objective— To (1) evaluate and compare computer-assisted surgery (CAS) with conventional screw insertion (conventional osteosynthesis [COS]) for treatment of equine abaxial distal phalanx fractures; (2) compare planned screw position with actual postoperative position; and (3) determine preferred screw insertion direction.
Study design— Experimental study.
Sample population— Cadaveric equine limbs (n=32).
Methods— In 8 specimens each, a 4.5 mm cortex bone screw was inserted in lag fashion in dorsopalmar (plantar) direction using CAS or COS. In 2 other groups of 8, the screws were inserted in opposite direction. Precision of CAS was determined by comparison of planned and actual screw position. Preferred screw direction was also assessed for CAS and COS.
Results— In 4 of 6 direct comparisons, screw positioning was significantly better with CAS. Results of precision analysis for screw position were similar to studies published in human medicine. None of evaluated criteria identified a preferred direction for screw insertion.
Conclusion— For abaxial fractures of the distal phalanx, superior precision in screw position is achieved with CAS technique compared with COS technique.
Clinical Relevance— Abaxial fractures of the distal phalanx lend themselves to computer-assisted implantation of 1 screw in a dorsopalmar (plantar) direction. Because of the complex anatomic relationships, and our results, we discourage use of COS technique for repair of this fracture type.