DNA metabarcoding provides great potential for species identification in complex samples such as food supplements and traditional medicines. Such a method would aid Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES) enforcement officers to combat wildlife crime by preventing illegal trade of endangered plant and animal species. The objective of this research was to develop a multi-locus DNA metabarcoding method for forensic wildlife species identification and to evaluate the applicability and reproducibility of this approach across different laboratories. A DNA metabarcoding method was developed that makes use of 12 DNA barcode markers that have demonstrated universal applicability across a wide range of plant and animal taxa and that facilitate the identification of species in samples containing degraded DNA. The DNA metabarcoding method was developed based on Illumina MiSeq amplicon sequencing of well-defined experimental mixtures, for which a bioinformatics pipeline with user-friendly web-interface was developed. The performance of the DNA metabarcoding method was assessed in an international validation trial by 16 laboratories, in which the method was found to be highly reproducible and sensitive enough to identify species present in a mixture at 1% dry weight content. The advanced multi-locus DNA metabarcoding method assessed in this study provides reliable and detailed data on the composition of complex food products, including information on the presence of CITES-listed species. The method can provide improved resolution for species identification, while verifying species with multiple DNA barcodes contributes to an enhanced quality assurance.

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The speciation of P in environmental samples is operationally defined, since it depends on the analytical method used. In this study, we compared four methods to measure P in solution: ion chromatography (IC), the malachite green colorimetric method (CM), the diffusive gradient in thin films technique (DGT) and, for total dissolved P, optical inductively coupled plasma (ICP). These methods were compared on three sets of solutions (filtered over <0.45 μm): solutions with model organic P compounds, suspensions of synthesized inorganic Fe and Al colloids loaded with P, and environmental samples. The environmentally relevant organic P compounds were only marginally detected by CM and IC. Substantial fractions of certain organic P compounds contributed to the DGT measurement. Colorimetric analysis of DGT eluates detected in general less P than ICP analysis, indicating that these organic P compounds sorbed on the zero sink layer. Phosphorus associated with inorganic colloids was completely recovered by CM, but not by IC and least by DGT. Measurements on a wide set of 271 environmental samples (soil pore waters, groundwaters, and surface waters) suggest that surface water P is largely present as orthophosphate and phosphate sorbed onto inorganic colloids, whereas organic P contributes more in groundwaters.

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