Pest risk analysis and invasive alien plants: progress through PRATIQUE*

The EPPO Decision‐support scheme (DSS) for Pest Risk Analysis (PRA) was fundamentally revised by the PRATIQUE EU project. Based on the examples of Polygonum perfoliatum (mile‐a‐minute‐weed) and Eichhornia crassipes (water hyacinth), its applicability for invasive alien plants was tested. By means of a comparison of the risk analysis procedures with EPPO schemes before and after the project, pest group specific enhancements are outlined. For the two plants tested here, documented invasion history as well as biotic and abiotic requirements show that they can establish and spread in Europe and cause damage. PRATIQUE revisions to the EPPO DSS increased transparency, user friendliness and consistency for PRAs on invasive alien plants. This was accomplished through more comprehensive rating guidance, and aspects such as allowing individual risk elements to be documented and summarized to give a conclusive end result.     

The raccoon (Procyon lotor) as potential rabies reservoir species in Germany: a risk assessment

Terrestrial wildlife rabies has been successfully eliminated from Germany predominantly as a result of the distribution of oral rabies vaccine baits. In case that wildlife rabies would re-emerge among its known reservoir species in Germany, swift action based on previous experiences could spatially and temporally limit and subsequently control such an outbreak. However, if rabies emerged in the raccoon population in Germany (Procyon lotor), there are no tools or local experience available to cope with this situation. This is especially worrisome for urban areas like Kassel (Hesse) due to the extremely high raccoon population density. A rabies outbreak among this potential reservoir host species in these urban settings could have a significant impact on public and animal health.     

Combination of TLC and HPLC-MS/MS methods. Approach to a rational quality control of Chinese star anise

In this study, a methodological approach for an effective and reliable quality control of Chinese star anise (Illicium verum Hook. F.) is developed and validated. A combined method of TLC and HPLC-MS/MS was used for differentiation of various Illicium species, especially Chinese and Japanese star anise. Species can be distinguished by their TLC flavonoid pattern. A sensitive and selective HPLC/ESI-MS/MS method was developed for the detection and quantification of lower admixtures of I. anisatum and of further toxic Illicium species at a low concentration range using the sesquiterpene lactone anisatin as a marker. The proposed assay includes a solid-phase extraction cleanup procedure with a high recovery (>90%). Chromatographic separation of anisatin was carried out on a C18 column, followed by MS detection using ESI in negative mode. The precursor/product ion transitions m/z 327 --> 127 (quantifier) and m/z 327 --> 297 (qualifier) were monitored. Statistical evaluation of this multireaction-monitoring procedure reveals good linearity and intra- and interday precision. The limits of detection and quantification are 1.2 and 3.9 microg/kg, respectively.     

Use of genetic profiling in leprosy to discriminate clinical forms of the disease

Leprosy presents as a clinical and immunological spectrum of disease. With the use of gene expression profiling, we observed that a distinction in gene expression correlates with and accurately classifies the clinical form of the disease. Genes belonging to the leukocyte immunoglobulin-like receptor (LIR) family were significantly up-regulated in lesions of lepromatous patients suffering from the disseminated form of the infection. In functional studies, LIR-7 suppressed innate host defense mechanisms by shifting monocyte production from interleukin-12 toward interleukin-10 and by blocking antimicrobial activity triggered by Toll-like receptors. Gene expression profiles may be useful in defining clinical forms of disease and providing insights into the regulation of immune responses to pathogens.     

Metal complexation by the peptide-bound maillard reaction products N(epsilon)-fructoselysine and N(epsilon)-carboxymethyllysine

Although the Maillard reaction between proteins and carbohydrates is of central importance for food processing and in vivo processes, only little is known about changes of the metal-binding properties induced by protein glycation. The purpose of this study was to examine the complex formation of the quantitatively important peptide-bound Maillard reaction products (MRPs) N(epsilon)-fructoselysine and N(epsilon)-carboxymethyllysine with the biologically relevant metal ions copper(II) and zinc(II). The MRPs were synthesized as the N(alpha)-hippuryllysine derivatives in order to block the coordination function of the alpha-amino group. Stability constant measurements were performed in aqueous solution using pH potentiometry. N(alpha)-Hippuryl-N(epsilon)-fructoselysine forms moderate Cu(II) complexes (Log(10) K(1) = 5.8; Log(10) K(2) = 4.0) but fails to form any complexes with Zn(II). N(alpha)-Hippuryl-N(epsilon)-carboxymethyllysine gives slightly stronger complexes with Cu(II) (Log(10) K(1) = 7.3; Log(10) K(2) = 6.3), but again no complexation with Zn(II) was observed. These results show that post-translational modification of proteins by carbohydrates leads to the formation of new coordination centers for metal ions within a protein chain. Further studies are necessary to clarify the consequences of this phenomenon in terms of protein quality and physiological processes.     

Quantitative impact of CO2 enriched atmosphere on abundances of methanotrophic bacteria in a meadow soil

In the Giessen free-air CO₂ enrichment (GiFACE) experiment, 5 years of CO₂ enrichment led to decreased CH₄ uptake rates of the investigated meadow soil. In soils, CH₄ is mainly oxidised by methanotrophic bacteria. In the present study, abundances of methanotrophic bacteria and total bacteria in soil samples from the GiFACE experiment were quantified by applying pmoA- and 16S rRNA gene-targeted real-time PCR and fluorescence in situ hybridisation (FISH). Methanotrophic bacteria of the Methylosinus group (Alphaproteobacteria) and the Methylobacter/Methylosarcina group (Gammaproteobacteria) were detectable by real-time PCR as well as by FISH. Both quantitative analytical approaches revealed that abundances of either bacteria or methanotrophic bacteria in soil samples from sites under CO₂-enriched atmosphere were decreased. Compared to ambient site, only 46 and 30.5% of methanotrophic bacteria and 38 and 63.2% of total bacterial cell numbers could be detected under CO₂-enriched atmosphere by FISH and real-time PCR, respectively. These results suggest that significantly decreased abundances of methanotrophic bacteria could explain reduced CH₄ uptake rates.