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Directive 2002/32/EC of the European Parliament and of the Council of 7 May 2002 on undesirable substances in animal feed lists a range of substances from botanical origin (weed seeds) and additionally some chemical compounds directly originating from specific weeds. In order to examine the actual status of enforcement and of the present occurrence of these botanical substances, a survey was carried out. A questionnaire was sent to 103 laboratories, including official control labs from all member states of the European Union. The results, indicating the frequency of occurrence as far as reported, are compared to the publications of the EU Rapid Alert System for Food and Feed (RASFF). A total of 44 questionnaires was returned (42.7%) from 22 member states. Ten member states predominantly from north-western Europe appeared to have an active monitoring of botanical undesirable substances. The questionnaire results did not indicate that the other member states enforce this part of Directive 2002/32/EC. Reports on the frequency of occurrence include: a few to 25-50% of the samples contain traces of ergot (8 member states), a few to 24% contain at least some traces of thorn apple (6 member states), zero to 17% contain some castor oil plant seeds (3 member states), zero to a few samples contain Crotalaria seeds (3 member states), and zero to 6% contain traces of Sareptian mustard (4 member states). One member state conducted extra surveillance since several cases of animal intoxications have been reported. In some cases a coincidence with undesirable botanical substances was found.

In the framework of the European Union funded research project STRATFEED for detection and identification of animal proteins in feeds, the microscopic method was optimized at several key steps and better documented. A check list was developed for uniform reporting. Characters of bone fragments, hairs, muscles and gills are fully documented. A so-called muscle ratio has been developed for the identification of muscle fibers at the level of vertebrate classes (mammals, birds and fishes). Both the improved protocol and the entire range of characters which can be observed, are documented in a Decision Support System called ARIES (Animal Remains Identification and Evaluation System). A second internet-based system called STRATFEED-DSS exclusively assists in identification of animal particles in feeds. A new strategy with microscopy as screening technique and either DNA or protein identification as confirmation technique is proposed. The advantages of this combination are the extremely low level of false negatives, low detection limits and the heat-resistant nature of microscopic detection, together with the possibility of a very specific identification of particles by one of the other methods.

Nontargeted 400 MHz (13)C and (1)H nuclear magnetic resonance (NMR) spectroscopy was used in the context of food surveillance to reveal Pinus species whose nuts cause taste disturbance following their consumption, the so-called pine nut syndrome (PNS). Using principal component analysis, three groups of pine nuts were distinguished. PNS-causing products were found in only one of the groups, which however also included some normal products. Sensory analysis was still required to confirm PNS, but NMR allowed the sorting of 53% of 57 samples, which belong to the two groups not containing PNS species. Furthermore, soft independent modeling of class analogy was able to classify the samples between the three groups. NMR spectroscopy was judged as suitable for the screening of pine nuts for PNS. This process may be advantageous as a means of importation control that will allow the identification of samples suitable for direct clearance and those that require further sensory analysis.

The bovine spongiform encephalopathy epidemic is thought to have occurred as a consequence of feeding prion-infected material to cattle. To avoid the risk of bovine spongiform encephalopathy diffusion, the European Commission (Directive 2003/126/EC) established an official method to detect the presence of animal-derived constituents in feedstuffs, using microscopic examination. This method allows easy identification of bone fragments among other animal constituents. The analysis is based on morphological conformation of the fragments and their characterization (mainly of the shape of lacunae) to discriminate among mammalian, poultry, and fish tissues. The aim of this study was to assess the performances of nine European laboratories through a ring trial of the official microscopic method, and to calculate accuracy and reproducibility of the method. In general the reproducibility of the microscopic method was very good (kappa overall = 0.83), with a high sensitivity for all laboratories. Concerning the analysis on the different animal-derived constituents, the results show values of sensitivity with large variability between fish and poultry or mammal. It was generally more difficult to discriminate between mammalian and poultry tissues than fish tissue.

In this paper we present an alternative method for detection of meat and bone meal (MBM) in feedstuffs by near-infrared microscopic (NIRM) analysis of the particles in the sediment fraction (dense fraction (d >1.62) from dichloroethylene) of compound feeds. To apply this method the particles of the sediment fraction are spread on a sample holder and presented to the NIR microscope. By using the pointer of the microscope the infrared beam is focussed on each particle and the NIR spectrum (1112-2500 nm) is collected. This method can be used to detect the presence of MBM at concentrations as low as 0.05% mass fraction. When results from the NIRM method were compared with the classical microscopic method, a coefficient of determination (R2) of 0.87 was obtained. The results of this study demonstrated that this method could be proposed as a complementary tool for the detection of banned MBM in feedstuffs by reinforcement of the monitoring of feeds.