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The tendency to achieve a circular bioeconomy and sustainability of the food production chain as far as possible has the consequence that an increasing variety of Former Food Products will be used as feed ingredient. The incidence of packed food stuffs in feed, and consequently unpacking upon use as feed ingredient, will increase when the by-products approach the final stages of processing. In this framework a method has been developed for examining the amount of packaging material in candy syrup, based on confectionery and sweets, with an approximate sugar content of 55%.

National Reference Laboratories (NRLs) are part of the system responsible for controlling and enforcing the EU food and feed law. Wageningen Food Safety Research (WFSR) has been designated the NRL for thirteen subjects. The tasks of an NRL depend on its research fields. This report gives an overview of the activities performed by all of WFSR’s NRLs in 2021. These NRLs are for: milk and milk products, marine biotoxins,animal proteins, certain substances and residues thereof as laid down in Regulation (EU) 2017/625, additives for use in animal nutrition (feed additives), genetically modified organisms (GMOs) in food and feed,pesticides, metals and nitrogenous substances in feed and food, mycotoxins and plant toxins in food and feed, processing contaminants halogenated persistent organic pollutants in food and feed, food-borne viruses and water content of poultry.This report first gives an overview of relevant legislation and information on the networks of EURLs, NRLsand OLs. For every NRL, a description of all activities performed in the EURL-NRL network, such as participation in EURL-NRL workshops, working groups, and proficiency and comparative tests. This is followed by a description of the assistance provided to OLs, such as a quality check or advice. Finally, the scientific and technical support given to the competent authority is discussed. In some cases, contact with other NRLs is discussed. An important NRL task is to stay updated with current developments within its NRL domain. Every EURL organises one or two meetings (workshops) every year for that purpose. Participation in these EURL-NRL workshops is mandatory. In 2021, because of COVID-19, the workshops were almost all held online. NRLs of WFSR have attended all workshops and actively participated in EURL working groups to improve analytical methods.To test the analytical capabilities of NRLs, the EURLs organise proficiency tests. As the scope of the EURL proficiency tests is sometimes limited, the NRLs al

WFSR report 2021.014 | 9 Summary National Reference Laboratories (NRLs) are part of the system responsible for the control and enforcement of EU food and feed law. Wageningen Food Safety Research (WFSR) has been designated as the NRL for thirteen subjects. The tasks of a NRL depend on its research fields. This report gives an overview of the activities performed by all of NRLs of WFSR in 2020. These NRLs are for: milk and milk products, marine biotoxins, animal proteins, certain substances and residues thereof as laid down in Directive 96/23/EC, additives for use in animal nutrition (feed additives), genetically modified organisms (GMOs) in food and feed, pesticides, metals and nitrogenous substances in feed and food, mycotoxins and plant toxins in food and feed, processing contaminants, halogenated persistent organic pollutants in food and feed, food borne viruses and water content of poultry. This report first gives an overview of relevant legislation and information on the networks of EURLs, NRLs and OLs. For every NRL, a description is then given of all activities performed in the EURL-NRL network such as participation in EURL-NRL workshops, working groups, and proficiency and comparative tests. This is followed by a description of the assistance given to OLs in the form of quality control and/or advice. Finally, the scientific and technical support given to the competent authority is discussed. In some cases, the contact with other NRLs is discussed. An important NRL task is to stay up to date with current developments within its NRL domain. Every EURL organises one or two meetings (workshops) every year for that purpose. Participation in these EURL-NRL workshops is mandatory. In 2020, due to COVID-19, most of these workshops were online. All workshops have been attended by NRLs of WFSR. Additionally, the NRLs have actively participated in EURL working groups, to improve analytical methods. To test the analytical capabilities of NRLs, the EURLs organise profici

National Reference Laboratories (NRLs) are part of the system responsible for the control andenforcement of EU food and feed law. Wageningen Food Safety Research (WFSR) has been designated as the NRL for twelve subjects. The tasks of a NRL depend on its research fields. This report gives an overview of the activities performed by all of WFSR’s NRLs in 2019. These NRLs are for: halogenated persistent organic pollutants in food and feed, pesticides in products of animal origin, mycotoxins and plant toxins in food and feed, metals and nitrogenous substances in feed and food, p rocessing contaminants, marine biotoxins, certain substances and residues thereof as laid down in Directive 96/23/EC, genetically modified organisms (GMOs) in food and feed, animal proteins, additives for use in animal nutrition (feed additives), milk and milk products, and water content of poultry.This report first gives an overview of relevant legislation and information on the networks of EURLs, NRLs and OLs. For every NRL, a description is then given of all activities performed in the EURL-NRL network such as participation in EURL-NRL workshops, working groups, and proficiency and comparative tests. This is followed by a description of the assistance given to OLs in the form of quality control and/or advice. Finally, the scientific and technical support given to the competentauthority is discussed. In some cases, the contact with other NRLs is discussed.An important NRL task is to stay up to date with current developments within its NRL domain. Every EURL organises one or two meetings (workshops) every year for that purpose. Participation in these EURL-NRL workshops is mandatory. In 2019, 1 5 workshops have been attended by NRLs of WFSR (former name, before June 2019, RIKILT Wageningen University & Research). Additionally, the NRLs have actively participated in EURL working groups to improve analytical methods. To test the analytical capabilities of NRLs, the EURLs organise proficiency tests.

A proficiency test (PT) was organised for the detection of packaging materials in bakery by-products intended to be used as feed ingredients. Two blind samples of a size of 250 grams at spike levels of 50 mg and 250 mg per sample were distributed to 29 participants. Four participants analysed only a part of the sample material, leaving 25 data points eligible for further evaluation. The evaluation of the results was carried out according to the principles of the Standard ISO 17043:2010 and the Quality Guidelines for visual research, in the framework of individual spiking of the samples. This specific procedure was chosen to avoid large inhomogeneity of a general batch as basis for the production of the PT samples, with the consequence that usual statistics such as Z-scores could not be applied or needed modified interpretations. Results of this PT have been compared with the first PT in 2016 (n=22 participants, same spike levels). Altogether this provided the presence of four datasets. Blanks were not included in both PTs. The average recovery for the 50 mg level was 179%, and 105% for the 250 mg level in the 2019 PT. The maximum overestimations were 441% (50 mg) and 170% (250 mg), respectively. Symmetrical recovery intervals were chosen of 34%-166% for the 50 mg level and 66%-134% for the 250 mg level. A share of 80% of the results for the 250 mg sample was achieved within the limits of the recovery interval. A considerable number of overestimations was reported for the 50 mg level (12 participants, 48%) in the 2019 PT. Notwithstanding the limitations of statistical parameters, a higher precision in 2019 compared to the 2016 datasets can be concluded based on the smaller interval of the results, visible in the minimum-maximum ranges, in the 95% confidence intervals and in the lower standard deviations. In all cases the distributions are skewed to the right, shown by the underestimations in only one dataset, lack of outliers below the Lower Confidence Level and skew

A proficiency test was organized for the microscopic determination and semi-quantification of botanic ingredients in the formulation of an animal feed, in the framework of the annual proficiency tests of the IAG - International Association for Feeding stuff Analysis, Section Feeding stuff Microscopy. The organizer of the proficiency test was Wageningen Food Safety Research, The Netherlands. The aim of the proficiency study was to provide the participants information on the performance of the local implementation of the method for composition analysis of feed. The current proficiency test was focusing on the control of the label declaration of the botanic composition of a ruminant feed. Participants received a sample of the ruminant feed and were asked to check the correctness of the label information. Therefore, the label declaration was provided together with the sample. The formulation as declared by the label showed the correct composition. Results should show the share of the different ingredients in percentages. Indicated shares were considered under- or overestimations when exceeding the limits of the IAG uncertainty interval model.A total of 22 sets of results was returned. Six of the nine ingredients had shares with intervals of which the lower limits were still higher than zero percent in the IAG estimation model. Correct estimation of these shares needs more precision than in a situation where only an upper limit applies. The reported shares of these six ingredients were within the limits of the uncertainty model in 87.9% of the total number of estimations. The correct estimations of all reports of the shares of nine ingredients is 80.3%. Six out of 22 participants delivered an errorless composition, which is 27%. Besides this, six participants made one error, three made two errors and two participants made three errors. There is no clear correlation with the method applied. Only three participants reported an indication of the correctness of the label. Th

National Reference Laboratories (NRLs) are part of the system responsible for the control and enforcement of EU food and feed law. RIKILT Wageningen University & Research has been designated as the NRL for twelve subjects. The tasks of a NRL depend on its research field. This report gives an overview of the activities performed by all of RIKILT's NRLs in 2016.

Fish meal is an accepted ingredient in compound feed. Unauthorised application is primarily enforced by visual inspection, i.e., microscopy. In order to document the visually available diversity, fragments of bones and scales of 17 teleost fish species belonging to seven different orders were investigated for their diversity in the presence of structural elements: lacunae and canaliculae in bone fragments and type of growth rings and teeth of scale fragments. Despite the classical division into cellular bones and acellular bones of teleost fish, i.e., whether or not possessing osteocytes, the current examinations revealed patterns of lacunae, in some types accompanied with canaliculae, in all 17 species investigated. In total seven types of bone structures were defined, and six types of scale structures. Profiles with the relative frequency of each bone type per species were established. The share of acellular bone fragments appeared to be related to the evolutionary position of the species. Results of proficiency tests for the detection of fish meal reveal that in most cases the sensitivity and specificity for the detection of fish meal ranges from sufficient to perfect. Only some specified circumstances can hamper proper recognition and identification, most notably salmon bone fragments mimicking bone fragments from terrestrial animals, and pieces of hydrolysed proteins or minerals mimicking acellular fish bone fragments. The expertise gained in this study would help to improve the distinction between fish meal and terrestrial animal material in compound feed, and it supports the application of the species-to-species ban with respect to the valorisation of by-products from fish farms in aquafeed. In a broader perspective, the current expertise might be helpful to detect fraud throughout the feed/food production chain. The matrix of characteristics versus species is implemented in a data model running in the expert system ‘Determinator’ for facilitating identificat

In the framework of sustainability and a circular economy, new ingredients for feed are desired and, to this end, initiatives for implementing such novel ingredients have been started. The initiatives include a range of different sources, of which insects are of particular interest. Within the European Union, generally, a new feed ingredient should comply with legal constraints in terms of ‘yes, provided that’ its safety commits to a range of legal limits for heavy metals, mycotoxins, pesticides, contaminants, pathogens etc. In the case of animal proteins, however, a second legal framework applies which is based on the principle ‘no, unless’. This legislation for eradicating transmissible spongiform encephalopathy consists of prohibitions with a set of derogations applying to specific situations. Insects are currently considered animal proteins. The use of insect proteins is a good case to illustrate this difference between a positive, although restricted, modus and a negative modus for allowing animal proteins. This overview presents aspects in the areas of legislation, feed safety, environmental issues, efficiency and detection of the identity of insects. Use of insects as an extra step in the feed production chain costs extra energy and this results in a higher footprint. A measure for energy conversion should be used to facilitate the comparison between production systems based on cold- versus warm-blooded animals. Added value can be found by applying new commodities for rearing, including but not limited to category 2 animal by-products, catering and household waste including meat, and manure. Furthermore, monitoring of a correct use of insects is one possible approach for label control, traceability and prevention of fraud. The link between legislation and enforcement is strong. A principle called WISE (Witful, Indicative, Societal demands, Enforceable) is launched for governing the relationship between the above-mentioned aspects.

The analysis of composition in terms of ingredients is important for detecting economic fraud and for monitoring feed safety. Composition analysis and label control of feed is regulated in Regulation (EC) 767/2009. In a broader view, composition analysis in the entire food chain can improve the effect of monitoring actions. The new legislation on food labelling (Regulation (EC) 1169/2011), effective from December 13th 2014, obliges to provide more detailed information to customers on composition and related topics. A ring test was organized for the microscopic determination of botanic composition in animal feed in the framework of the annual ring tests of the IAG - International Association for Feeding stuff Analysis, Section Feeding stuff Microscopy. The organizer of the ring test was RIKILT - Wageningen UR, The Netherlands. The aim of the ring study was to provide the participants information on the performance of the local implementation of the method for composition analysis of feed. The sample was based on an artificially produced feed mimicking a ruminant feed, and distributed without label information. The participants were requested to produce a correct declaration of the ingredients of the sample. The results were analysed using the IAG model for uncertainty limits. Shares of ingredients in the feed formulation outside the limits of the model were indicated as underor over-estimations. A total of 25 sets of results was returned. The percentage of under- or over-estimations was 28.6% for the seven main ingredients. In the overview of results, the two declared wheat ingredients and the two declared corn products were pooled to one ingredient each. This was necessary since some participants declared a general ingredient (“wheat” and “corn”) and others a specific type (gluten or bran). The use of the original declarations would result in an extra number of non-matching estimations without precise justification. The share of the citrus pulp, in the presence of a

The annual ring test for the detection of animal proteins in animal feed of the IAG - International Association for Feeding stuff Analysis, Section Feeding stuff Microscopy was organized by RIKILT - Wageningen UR, The Netherlands. The aim of the ring study was to provide the participants information on the performance of the local implementation of the detection method for their local quality systems. A further aim was to gather information about the application of the microscopic method. The current 2016 version of the IAG ring test for animal proteins facilitated the full scenario with the methods for microscopy and PCR as published in Regulation (EC) 51/2013 amending Annex VI of Regulation (EC) 152/2009 together with accompanying SOPs. All four samples were based on an artificial feed mimicking a formulation for ruminant feed. Two samples were labelled as fish feed (B and D), which was effectuated by adding 2% of a general fish meal. Adulteration was achieved by adding 0.1% pig MBM (B), 0.1% ruminant MBM (D) and a combination of 0.1% ruminant MBM and 0.1% fish meal (C). This combination of different spikes allowed the diverse application of the detection methods. Forty eight participants enrolled for the ring test, of which 45 submitted microscopic results. Of these, 20 participants applied the combination of microscopic and PCR analysis. Three participants submitted exclusively PCR results.

Ergot alkaloids are recognised as seriously toxic compounds, which caused a series of outbreaks in the past. In the EU, enforcement is implemented by visual detection and quantification of ergot sclerotia produced by moulds of the genus Claviceps. On behalf of the IAG section Feedstuff Microscopy, RIKILT organised a ring test for the visual detection of ergot sclerotia in two unground rye samples in September 2015. In this report the results from the ring test for ergot in rye 2015 are presented. The ring test ergot sclerotia in rye was designed to test the capability to visually detect sclerotia or parts thereof at relatively high levels. One sample was based on a level of approx. 400 ppm, and the second sample contained an amount of approx. 1000 ppm (EU legal limit for feeds and ingredients: 1000 ppm = 1 gram/kg = 0.1%). An amount of approx. 250 grams of rye grains was chosen as sample size. All samples were individually spiked. Thirty participants enrolled for the ring test. Participants were requested to report the number of recovered (fragments of) sclerotia and the total weight per sample. The percentage of recovery for every sample was calculated. A dedicated IAG method as well as other (lab internal) methods were allowed for application. Principally, methods are based on sieving (preferably with a mesh size of 0.5 mm), examination of every particle (grain) in the fraction with full grains or particles larger than 0.5 mm, selection of sclerotia fragments supported by documentation, and weighing the final selection of bodies. The average recovery for both samples was approx. 97%. All results except one were between the expected recovery limits (80 – 110 % w/w). Supporting data from an intralaboratory validation study of the IAG method showed trueness at different low spike levels between 98 and 105% w/w. Limit of detection was established at 7 ppm. It can be concluded that examination by visual detection of sclerotia is a valuable indicator of the expected pre