Mayflies, stoneflies and caddisflies (Ephemeroptera, Plecoptera and Trichoptera) are prominent representatives of aquatic macroinvertebrates, commonly used as indicator organisms for water quality and ecosystem assessments. However, unambiguous morphological identification of EPT species, especially their immature life stages, is a challenging, yet fundamental task. A comprehensive DNA barcode library based upon taxonomically well-curated specimens is needed to overcome the problematic identification. Once available, this library will support the implementation of fast, cost-efficient and reliable DNA-based identifications and assessments of ecological status. This study represents a major step towards a DNA barcode reference library as it covers for two-thirds of Germany's EPT species including 2,613 individuals belonging to 363 identified species. As such, it provides coverage for 38 of 44 families (86%) and practically all major bioindicator species. DNA barcode compliant sequences (≥500 bp) were recovered from 98.74% of the analysed specimens. Whereas most species (325, i.e., 89.53%) were unambiguously assigned to a single Barcode Index Number (BIN) by its COI sequence, 38 species (18 Ephemeroptera, nine Plecoptera and 11 Trichoptera) were assigned to a total of 89 BINs. Most of these additional BINs formed nearest neighbour clusters, reflecting the discrimination of geographical subclades of a currently recognized species. BIN sharing was uncommon, involving only two species pairs of Ephemeroptera. Interestingly, both maximum pairwise and nearest neighbour distances were substantially higher for Ephemeroptera compared to Plecoptera and Trichoptera, possibly indicating older speciation events, stronger positive selection or faster rate of molecular evolution.
In soils, colloidal transport has been identified as the most important pathway for strong adsorbing, environmental contaminants like pesticides, heavy metals, and phosphorus. We conducted a comparative dye tracer experiment using a Brilliant Blue (BB) solution and a Titanium(IV) oxide (TiO 2 ) colloid suspension (average particle size 0.3 mm), aiming to visualize and quantify colloid pathways in soils. Both dye tracers showed comparable general flow patterns with preferred transport over the deepest part of the soil profile, independent of clay content. The stained area was generally smaller for TiO 2 than for BB by a factor of ten, however, and there was no TiO 2 to be found at all in the low clay content soil. The travel distance was almost identical for the solution and the suspension (0.7 m) giving evidence that environmentally critical compounds bound to microparticles may be vertically transported over longer distances in soils, even within single rainfall events. The spatial variability of the dye patterns was large on a small scale with a range of 0.35 m for TiO 2 in the horizontal plane, which was taken as a general proof for a pronounced preferential transport situation. The study indicates that TiO 2 is transported exclusively through singular macropores of biogenetic nature, while BB passes also through the soil matrix of coarse-bedded soils, the secondary pore system or interaggregate pore space. The results emphasize the general suitability of TiO 2 for the visualization of colloid transport pathways in soils, opening up new research opportunities for contaminant transport in soils.
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