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.     

Climatic response and impacts of drought on oaks at urban and forest sites

This study focuses on the climate and growth response of the sessile oak (Quercus petraea (Matt.) Liebl.) and the northern red oak (Quercus rubra L.) using dendrochronological methods. Tree-ring series of street trees in the city of Dresden, Germany and in a nature reserve close to the urban area were analysed, and the climate and growth response and moving correlation functions (CF) were computed. Moreover, the impact of 13 years of drought was examined by superposed epoch analysis (SEA). Our results show that differences emerge from peculiarities of the different site conditions, indicating that city oaks contain a higher level of noise in their ring width data due to human activities that degrade the strength of growth-climate associations. Importantly, climatic conditions during the previous summer and autumn and during the current spring season display significant correlations with annual growth. The stability of the climate–growth relationship was tested via moving correlation functions. A strong increase in the correlation with the temperature and precipitation in April was found for the trees of northern red oak in forests. Drought years induced significant reductions in radial growth for the northern red oak only. Nevertheless, the long-term impact of drought years must be considered as marginal because of the fast recovery of the radial growth after one to two years. In the future, the growth of the northern red oak may be restricted by an increased frequency of droughts and warmer and drier conditions in April.     

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.     

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.     

Contamination of highly weathered urban soils in Uberlândia,Brazil

We determined heavy metal, polycyclic aromatic hydrocarbon (PAH), and polychlorinated biphenyl (PCB) concentrations in 18 topsoils of Uberlândia (420,000 inhabitants, Brazil) and in 3 rural topsoils. Concentrations of Al (11—124 g kg^—1) and Fe (13—109 g kg^—1) are large because of desilification. Concentrations of Cd (0.1—0.5 mg kg^—1), Cr (13—72), Cu (6—154), Mn (28—974), Ni (4—29), Pb (3—26), Zn (4—107), the sum of 20 PAHs (=Σ20PAHs:7—390 μg kg^—1), and the sum of 14 PCBs (=Σ14PCBs:0.05—1.25) are comparable to or below background concentrations in temperate soils except for Cu at two sites. More than 67% of the metals are strongly bound in Fe oxides and silicates; metals are more bioavailable in the urban than in the rural soils. The most abundant PAHs in the urban soils, on average, are naphthalene (19.0 ± 13.4% of Σ20PAHs) and the benzo(b+j+k)fluoranthenes (11.4 ± 6.7%); the most abundant PCBs are nos. 138 (23.3 ± 11.0% of Σ14PCBs) and 153 (14.3 ± 6.4%). The rural soils contain larger percentages of low molecular PAHs and up to tetra‒chlorinated PCBs than the urban soils. The different pollutant concentrations and patterns in the studied tropical compared with many temperate soils indicate different sources and fate.     

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.