feed 2016feedsafety.org/wp-content/uploads/2014/02/programme-book2016.pdf · 2 general introduction...

19 - 20 October 2016 Geel - Belgium

Feed 20165th International Feed Conference:Present and Future Challenges

Programme & Abstract book

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General Introduction

On behalf of the European Commission’s Joint Research Centre (JRC) it is my honour to welcome you to the 5th International Feed Conference 2016 at the JRC-Geel site. The availability of sufficient and safe feedingstuff is a key challenge in modern agriculture. Since the beginning of this millennium, the JRC has been working together with Member States’ expert laboratories on various topics related to feedingstuffs, including giving support to manage the crises related to BSE and dioxins. The JRC also undertook actions to make processed animal proteins safe to be returned to the feeding stuff sector, all of this leading to significant improvement of the entire system of compliance check with legal requirements. While the challenge from the undesirable substance in feed remains a major issue, the globalisation of the feed business has further reinforced the need for efficient tools for traceability of feed ingredients. Moreover, the constantly increasing demand for food from the animal origin, along with limited resources triggers the need for evaluating new sources of feed ingredients such as insects. Keeping the feeding safe therefore requires a multidisciplinary approach, bringing together all stakeholders, including the industry. With the four entirely different sessions of this International Feed Conference, the organisers tried to reflect this complexity and the scientific programme includes very interesting contributions to these topics. To this end, I would like to wish you a successful meeting, fruitful discussions and an excellent stay at the JRC site Geel. E. AnklamDirector of JRC-Directorate F (Health, Consumers & Reference Materials)Directorate General Joint Research CentreEuropean Commission

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Organizing Committee

A. Boix (chair) U. Vincent (chair) C. von Holst (chair)

V. Baeten S. Bellorini M. Chedin J. de Jong Z. Ezerskis J. Fernández Pierna

M. J. González de la Huebra J. Omar Onaindia P. Robouch F. Serano B. Slowikowski L. van Raamdonk

Scientific Committee

V. Baeten, Belgium A. Boix, Belgium J. Brufau, Spain R. Codony, Spain J. de Jong, The Netherlands J. A. Fernández Pierna, Belgium C. Genouel, France L. Han, China L.A.P. Hoogenboom, The Netherlands

A. Karus, Estonia L. Pinotti, Italy P. Robouch, BelgiumJ. Sloth, Denmark L.W.D. van Raamsdonk, The Netherlands P. Veys, Belgium U. Vincent, Belgium C. von Holst, Belgium S. Weigel, Germany

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Keynote Speakers

Opening address: Sabine Kruse

Sabine Kruse holds a PhD in agriculture economics from the Humboldt University Berlin. She worked as a scientist at the Institute of Agricultural Economics in Berlin from 1979 to 1990. In 1991 she moved to the German Federal Ministry of Food and Agriculture, first in the business economics unit and since 1996 in the animal nutrition unit.Dr. Kruse is responsible for legislation relating to feed safety and animal nutrition in Germany. In particular she deals with all matters relating to undesirable substances, additives, feed hygiene and labelling. She also coordinates the feed control in Germany. She has been the head of the German Delegation to the Standing Committee for Animal Nutrition by the European Commission. Finally she has been the head of the German Delegation to the Codex Alimentarius Task Force of Animal Feeding.

Session 1: Avo Karus

D.Agr Avo Karus is professor of animal biochemistry in department of food science and food technology of Institute for Veterinary Medicine and Animal Science of the Estonian University of Life Science since 1997. He graduated in Chemistry at the University of Tartu. In 1990 gained a PhD in Biochemistry at the K.I.Skryabin Moscow State Academy of Veterinary Medicine and Biotechnology, Russia. In 1995 him was awarded Doctor of Agriculture (in Animal Sciences) degree at the Estonian Agricultural University. He is also a national expert at the Horizon 2020 Program Committee: Societal Challenge 2 Food security, sustainable agriculture and forestry, marine, maritime and inland waterway studies and the bio-economy. Professor Karus participated in several international projects and has contributed as external expert, rapporteur and evaluator of numerous Projects, Actions and Institutions.

Session 2: Paul Brereton

Paul Brereton is Head of Agri-food Research at Fera based in York, UK. As senior scientist working at science-policy interface, he has over 30 years’ experience of applied research in the area of food safety and quality. As well as leading numerous

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national activities, Paul has co-ordinated two of the world’s largest research projects in the area of food authenticity and traceability: the €20M TRACE project (Tracing the origin of food) and currently FOODINTEGRITY a €12M EU sponsored research project that aims to address, at a European level, many of the gaps identified during the recent horsemeat incident. He has published over 70 peer reviewed papers on food safety and quality and recently edited the book “New analytical approaches for verifying the origin of food” (Woodhead/Elsevier). He has close links with the food industry, UK Public sector, academia and the European Commission. In addition to his Fera role, he has also taken up a position at the Institute of Agri-Food Research and Innovation (IAFRI) a joint Institute between University of Newcastle and Fera that aims to deliver innovation in research across the agri-food chain.

Session 3: Frans Verstraete

Frans Verstraete graduated in 1985 as agricultural engineer at the University of Ghent (Belgium). After his studies he held positions at the University of Ghent and thereafter at the Belgian Ministry of Agriculture and he was for a period technical adviser of the Belgian Minister of Agriculture. He is working for the European Commission since 1997. In the European Commission he has had various functions but since 2000 he is working at the Directorate General Health and Food Safety. He is responsible for the elaboration, development and management of the EU-legislation concerning certain contaminants in feed and food.

Session 4: Peter Radewahn

Peter Radewahn Peter Radewahn had his education at the University of Bonn from 01.10.1977 until 30.04.1983 with several interruptions for practical studies and education on farm. He completed his university education with an academic degree: Diplom-Ingenieur for Agriculture which could be compared to M.Sc. today. Peter got his first employment as scientific assistant in Bundesverband der Mischfutterhersteller (Federal Association of Compound Feed Manufacturers) in Bonn on 01.07.1983. Then, since 1994, he had been the Managing Director of this Federal Association in Bonn. In 2000 this Federal Association of Compound Feed Manufacturers merged with another feed association resulting in the Deutscher Verband Tiernahrung e. V. (DVT) from which Peter Radewahn is also the Managing Director.

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Programme

Wednesday 19 October 2016

8:15-9:00 Registration

9:00-9:30 Opening session of the Feed 2016 Conference: Elke Anklam, Directorate General Joint Research Centre, Director of Directorate F - Health, Consumers & Reference Materials

Opening address: OutcomeoftheJointWHO/FAOexpertmeetingonhazardsin

feed 2015 Sabine Kruse, German Federal Ministry of Food, Agriculture and

Consumer Protection (BMEL), Germany

Session 1: New feed materials and security (availability of feed/food)

Chairs: Christoph von Holst, DG JRC European Commission & Vincent Baeten, CRA-W

9:30-10:00 Keynote speaker 1: Avo Karus (Estonian University of Life Sciences, Institute of Medicine & Animal Science, Estonia)

Valorizationoffoodby-productsandwaste-challengesandopportunities

10:00-10:20 Marta Ponghellini (O1) - DG SANTE, European Commission UpdateonFeedAuthorisations:NewAuthorisations:New

challengesandinnovationinFeedAdditivesector

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10:20-10:40 Matteo Ottoboni (O2) - Department of Health, Animal Science and Food Safety VESPA, Università degli Studi di Milano, Milan, Italy

Lightmicroscopytechniqueforthediscriminationofinsectprocessedanimalproteinsversusmarinearthropods

10:40-11:00 Marc Berntssen (O3) - National Institute of Nutrition and Seafood Research, Nordnes, Bergen, Norway

Modellingthefeed-to-foodtransferofpharmaceuticalsinnovelAtlanticsalmonfeedsbasedonprocessedanimalproteins(PAPs)

11:00-11:30 Coffeebreak,posterviewing

11:30-11:50 Rafael Codony (O4) - LiBiFOOD Research Group; Nutrition, Food Science and Gastronomy Department, University of Barcelona, Barcelona, Spain

Oilstandardization:thekeysteptobedonetoencouragetheuseofacidoils?

11:50-12:10 Luciano Pinotti (O5) - Department of Health, Animal Science and Food Safety VESPA, Università degli Studi di Milano, Milan, Italy

NutritionalevaluationofFormerFoodProductsintendedforpignutrition

12:10-12:30 Leo van Raamsdonk (O6) - RIKILT-Wageningen University and Research Centre, Wageningen, The Netherlands

Newfeedingredients:theinsectopportunity

12:30-14:00 Lunch and poster viewing

14:00-14:30 Poster session 1


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Session 2: Traceability/authenticityoffeedmaterialsincludingadditives

Chairs: Pierre Dardenne, CRA-W & Ana Boix, DG JRC European Commission

14:30-15:00 Keynote lecture 2: Paul Brereton - Food and Environment Research Agency (FERA), United Kingdom

Newsystemsforidentifyingpotentialrisksinthefeedchain

15:00-15:20 Jens Sloth (O7) - Technical University of Denmark, National Food Institute (DTU Food), Søborg, Denmark

Determinationofiodineandinorganicarsenicinfeed:analyticalmethodsandriskassessment

15:20-15:50 Wulf-Dieter Moll (O8) - BIOMIN Research Center, Tulln, Austria OligonucleotidesofDNAasspecific,biologicaltracers

15:50-16:10 Cathal Connolly (O9) - Alltech European Bioscience Centre, Sarney, Summerhill Road, Dunboyne, Co. Meath, Ireland

Theuseofspectroscopyinthefieldofmineralchelateanalysis

16:10-16:40 Tea break & poster viewing

16:40-17:00 Zengling Yang (O10) - College of Engineering, China Agricultural University, Beijing, P.R. China

Traceabilityandauthenticityoffeedmaterialsusingnearinfraredspectroscopyandnearinfraredmicrospectroscopy:casestudies

17:00-17:20 Ana Boix (O11) - DG JRC European Commission SpectroscopyappliedtofeedadditivesoftheEuropeanReference

Laboratory:animportanttoolfortraceability

17:20-18:00 Poster session 2

18:00 shuttle to hotels

19:30 Conference dinner

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Thursday 20 October 2016

Session 3: Contaminants,undesirablesubstances,residues

Chair: Jacob de Jong, RIKILT & Ursula Vincent, DG JRC European Commission

9:00-9:30 Keynote lecture 3: Frans Verstraete - DG SANTE, European Commission EU

Policyonundesirablesubstancesinfeed:recentdevelopmentsandoutlook

9:30-9:50 Monica Sanden (O12) - National Institute of Nutrition and Seafood Research (NIFES), Bergen, Norway

Levelsofundesirablesubstancesinmarinefeedingredients,commercialNorwegianfishfeedandfarmedAtlanticsalmoninthelast13-15years

9:50-10:10 Olivier Fumière (O13) - Walloon agricultural Research centre (CRA-W), Agricultural Product Valorisation Department/European Union Reference Laboratory for Animal Proteins in Feedingstuffs (EURL-AP), Gembloux, Belgium

Validationofaselectedpigreal-timePCRassayforthedetectionofprocessedanimalproteinsinfeedingstuffs

10:10-10:30 Ursula Vincent (O14) - DG JRC European Commission Candidatestandardmethodforthedeterminationofauthorised

coccidiostatsandbannedanti-bacterialsubstancesinanimalfeedingstuffs

10:30-11:00 Coffeebreakandposterviewing

11:00-11:20 Jacob de Jong (O15) - RIKILT-Wageningen University and Research Centre, Wageningen, The Netherlands

Ananalyticalstrategyfortheearlyqualityandsafetyassuranceintheglobalfeedchain


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11:20-11:50 Erik Nordkvist (O16) - National Veterinary Institute, Uppsala, Sweden

Naturalchloramphenicolincerealstraw–occurrence,transfertopigsandpossibleimplicationsinofficialcontrol

11:50-12:10 Jorge Numata (O17) - Federal Institute for Risk Assessment (BfR), Berlin, Germany

ComputationaltoxicokineticmodelingforconsumerprotectionagainstcontaminantsinfeedandfoodattheFederalInstituteforRisk Assessment (BfR)

12:10-13:30 Lunch and poster viewing

Session 4 Theindustryperspective

Chair: Lujia Han, CAU & Joerg Seifert, FEFANA

13:30-14:00 Keynote lecture 4: Peter Radewahn - Deutscher Verband Tiernahrung (DVT), Germany & FEFAC Belgium

14:00-14:20 Frank Gort (O18) - SecureFeed, Wageningen, The Netherlands SecureFeed–acollectivefeedsafetyapproachtomeetchanging

demandsinthefoodchain

14:20-14:50 Carine van Vuure (O19) - Darling Ingredients International, The Netherlands & EFPRA Belgium

UtilisingAnimalBy-Productsasasustainablefeedchoiceforfoodproducinganimals

14:50-15:10 Vincent Baeten (O20) - Walloon agricultural Research centre (CRA-W), Gembloux, Belgium

Analysis of feed at farm and industry levels

15:10-15:40 Tea break and poster viewing

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15:40-16:00 David Eisenberg (O21) - Anresco Laboratories, San Francisco, The United States

PMicrotracers(R)andtheiruseincodingfeedproductsandpremixesinfinalfeedstoassurefeedqualityandtoassureauthenticity

16:00-16:20 Yvan Dejaegher (O22) - BEMEFA-APFACA, Brussels, Belgium FeedSafety:Regional,nationalandinternationalinitiatives. AChallengeforthefuture

16:20-16:50 Closingsession–ChristophvonHolst,DGJRCEuropeanCommission

17:00 shuttles

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Abstracts

Opening Address:

OUTCOMEOFTHEJOINTWHO/FAOEXPERTMEETINGONHAZARDSINFEED2015

Kruse S. Unit feed safety and animal nutrition, Federal Ministry of Food and Agriculture, Bonn, Germany

In 2015, the FAO and the WHO invited 17 experts from 14 countries to the Joint FAO/WHO Expert Meeting on Hazards Associated with Animal Feed. The aim was to deliver an update on the state of knowledge on hazards regarding feed. The purpose of this presentation is to summarise the key conclusions of the meeting.Potential hazards: The expert meeting reviewed and discussed potential hazards in feed of chemical, biological and physical origin. For each of the hazards, the experts described the health impact, its source, the occurrence in feed, the transfer from feed to food and the relevance for food safety.

· As chemical hazards in feed, the expert meeting identified and assessed:· Persistent organic pollutants, such as dioxins (PCDDs), furans (PCDFs), dl-PCBs and

ndl PCBs · Veterinary drug residues · Residues of organochlorine and other pesticides· Toxic elements (e.g. arsenic, cadmium, lead, mercury) · Mycotoxins · Plant toxins (e.g. pyrrolizidine alkaloids, glucosinolates)· Other potential chemical hazards (e.g. brominated flame retardants, perfluorinated

compounds, chloroparaffins)

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As biological hazards in feed, the expert meeting identified and assessed: · Bacteria (Salmonella, Mycobacterium, Brucella, Clostridium, Escherichia coli,

Listeria)· Parasites · Viruses· Prions

As physical hazards in feed, the expert meeting identified and assessed: · Radionuclides· Residues of nanomaterials· Micro- and nano-plastics · other relevant materials (e.g. residues from packaging)

New feed materials: The expert meeting defined and analysed the hazards of feed and products of feed production technologies of increasing relevance, such as

· Insects as feed· Food waste and former food products e· Biofuel wastes· Aquatic plants· Marine resources

Such “new” feed materials present new challenges for risk assessment and management.The expert group gave various recommendations, such as the need for screening and confirmatory methods suitable for the assessment of the risks as mentioned above. Moreover, the main stake holders in the feed sector are encouraged to launch different activities like developing guidelines in the various fields, conducting research on risk assessment and organising specific training workshops on risk assessment and management of hazards in feed.


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Session 1: New feed materials and security (availability of feed/food)

Keynote 1: Valorizationoffoodby-productsandwaste–challengesandopportunities

Karus A. Estonian University of Life Sciences, Institute of Veterinary Medicine & Animal Science, Estonia

To meet the food demand to feed a global population of over 9 billion in 2050, a 60% increase in food production is needed in comparison with 2005 as declared in UN World Food Programme (2012). This can only be possible together with a drastic reduction of the huge losses and waste of food, and by a sustainable management of our natural resources. As fully recognised, food production is here competing with feed industry and bio-energetics for the available resources. Thus, the main driver for future development in food and feed primary sector is resource efficiency. To increase the efficiency, there are three mainstreams – to reuse by-products and waste for novel food production, to produce feed, or to use them for technical (often to produce energy or fertilizers) purposes. The greatest importance have animal origin products (and animal origin proteins in particular) because the production of animal origin proteins is costly itself. Overall one third of animal origin proteins will be wasted and 15 million tons of processed animal origin proteins will found in animal origin by-products per year worldwide. Proteins of animal origin should comprise approximately half of the total protein. The challenges in European scientific community and stockholders opinions are often related to concerns about quality and safety. In some areas as non-ruminant processed animal protein in fish feed in the EU and pet food are some examples of recognition of animal origin proteins in feed. In Europe, the annual pet food production is over 8 million tons with the annual growth rate 2.0% (average value over 3 years) and represents a combined annual turnover of over € 24 billion. To continue this trend in other fields of feed industry a proper risk assessment is needed and all decisions should be made based on reliable scientific evidence. The European possible developments, issues and public concerns as well as some bottom-up scientific actions will be discussed.

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O1. UpdateonFeedAuthorisations:NewAuthorisations:NewchallengesandinnovationinFeedAdditivesector.

PonghelliniM. European Commission, Directorate General SANTE, Brussels, BelgiumIn the recent past and at present, the sector has had to confront to new challenges not only on animal or human and environmental safety, but also on a deep economic crisis. The industry was sought to develop new products to address the aspects of Antimicrobial Resistance (AMR), animal welfare or new purposes on zootechnical effects. Therefore, in line with the new objectives of the Commission on the development and innovation, the legislator proposed some legislative initiatives aimed to encourage the development of new products, by amending either the Regulation (EC) 1831/2003 introducing new functional groups, either Regulation (EC) No 429/2008 by the introduction of new rules for the submission of dossiers, highlighting the new objectives in the field of animal welfare and productivity, without neglecting the general safety requirements.

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O2. Lightmicroscopytechniqueforthediscriminationofinsectprocessedanimal proteins versus marine arthropods

OttoboniM.1, Tretola M.1, Caprarulo V.1, Marchis D.2,VeysP.3, Baeten V.3,PinottiL.1 1 Department of Health, Animal Science and Food Safety VESPA, Università degli Studi di Milano, Milano, Italy2 Cento di Referenza per la Sorveglianza e il Controllo degli Alimenti per Animali C.Re.A.A, Istituto Zooprofilattico Sperimentale di Piemonte, Liguria e Valle D’Aosta, Italy3 Food and Feed Quality Unit, Valorisation Department, Walloon Agricultural Research Centre CRA-W, Gembloux, BelgiumThe new Novel Food Regulation has recently included whole insects under the scope of its application. However, the EU still needs to resolve a number of remaining legal questions in order to guarantee food and feed safety. In this context, it is important to develop a robust analytical method for the detection of insect originated material in animal feed. Starting from the assumption that insect meals, when authorized for feed formulation, will be considered as processed animal proteins (PAPs), the existing methods for PAPs identification (i.e. light microscopy and PCR) represents a robust starting point.

Accordingly, this study evaluated the use of light microscopy for the discrimination of insect PAPs against marine arthropods – classified as fish meal. Specifically, two samples of single


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species insect material, Hermetia Illucens (HI) and Tenebrio molitor (TM) and two samples of marine arthropods, shrimp material and krill, have been analysed and compared after staining by two reagents to enhance fragments identification. Alizarin Red (AR) and Chlorazol Black (CB), which react respectively with calcium salts and chitin, were tested for their potential efficacy in distinguish between insect and marine materials. Results obtained indicated that AR failed at staining HI and TM materials. By contrast, both HI and TM materials were stained by CB. When shrimp fragments and krill were considered, AR and CB stain tests colored marine materials reddish-pink and dark black, respectively. By combining these results it can be suggested that CB staining could be efficiently used to mark insect materials, AR does stain shrimp fragments but did not stain the tested insect material, indicating a possible approach for discriminating between insects and marine arthropods. However, since the present study has only been done on pure materials and on a small set of samples, possible implementation of these staining still need to be confirmed in complex matrices such as compound feed.

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O3. Modelingthefeed-to-foodtransferofpharmaceuticalsinnovelAtlanticsalmon feeds based on processed animal proteins (PAPs)

Berntssen M. H.G.1,Nácher-MestreJ.2,3, Bokkers B.4, Zeilmaker M.J.4

1 National Institute of Nutrition and Seafood Research, Bergen, Norway2 Research Institute for Pesticides and Water (IUPA). Avda. Sos Baynat, s/n. University Jaume I, Castellón, Spain3 Institute of Aquaculture of Torre la Sal IATS, CSIC, Castellón, Spain4 National Institute of Public Health and the Environment, Bilthoven, the Netherlands

The use of processed animal proteins (PAPs) such as feather meal, poultry by-product meal, pork meat, and poultry and pork blood meal have been shown to be relevant nutritional replacements of fish meal for many cultured fish species, including salmonids. Following a bovine spongiform encephalopathies (BSE) risk assessment by the European Food Safety Authorities (EFSA), in 2013 the EU set out a working plan for the re-authorization of the use of non-ruminant PAPs in animal feeds, initially for aquafeeds. These PAPs can harbor undesirable substances such as pharmaceuticals and their metabolites which are not previously associated with salmon farming, and might cause a potential risk for feed and food safety (Nacher-Mestre et al. 2016). These include authorized pharmaceutical products such as trimethoprim for which the EU has set MRLs in fish products, as well as none-permitted residues such as the dyes leucocrystal violet and leucomalachite green. Whether the residues will pose a risk to food safety will depend on their feed-to-fillet transfer. In the present study,

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a serial one-compartment physiological kinetic model, that includes differentiated growth of body components, is used to analyze the fillet accumulation of the pharmaceuticals in Atlantic salmon. The model is assessed from experimental uptake and elimination kinetics of Atlantic salmon fed the residues at two levels for 40 days followed by a depuration period of 90 days.

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O4. Oilstandardization:thekeysteptobedonetoencouragetheuseofacidoils?

CodonyR. 1, Barroeta A.C. 2, Sala R.2, Tres A.1, Varona E.1,GuardiolaF.11 LiBiFOOD Research Group, University of Barcelona, Barcelona, Spain 2 Departament de Ciència Animal i dels Aliments. Facultat de Veterinària, Universitat Spain

Autònoma de Barcelona, Bellaterra, SpainAcid oils are a by-product of oil refining, rich in free fatty acids (FFA), usually cheaper than crude or refined oils. However, in the 80s, low digestibility values were reported in chickens and pigs and this was attributed to their high FFA value, which did not favor their widespread use for feed uses. In order to valorize them, new technical fats obtained from the chemical esterification of acid oils with glycerol have been assayed as feed ingredients in our recent studies. The hypothesis was that the costs from the chemical esterification would be compensated by an increase in fat digestibility as a consequence of the FFA reduction. Moreover, the re-esterification process could be designed to obtain triacylglycerols together with a blend of di- and monoacylglycerols that could exert an emulsifying role during digestion. But our results in chickens and pigs have shown that these partially re-esterified fats have null or just a slight advantage in animal growth or product composition over the use of crude or acid oils. Therefore, re-esterified fats would only be economically advantageous when the costs of the re-esterification processes would be compensated by the productive benefits.

Moreover, the results obtained for acid oils were not always as detrimental as reported by other studies. However, we noticed a great variability in the composition of acid oils. This is one of the reasons why farmers refuse to use them although being cheaper fats. Also, the oxidative stability and shelf life of feed with added acid oils could become a concern. Nowadays, our research efforts are in the direction of completely characterizing acid oils available in the market to detect the main variability parameters affecting feed shelf life, animal production and the composition and stability of animal food products, so that recommendations on their use and control programs can be established. The presence of persistent organic pollutants in these acid oils also deserves attention.

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O5. NutritionalevaluationofFormerFoodProductsintendedforpignutrition

Giromini C., Tretola M.,OttoboniM.,GottardoD.,CastricaM.,CapraruloV.,CheliF.,PinottiL.Università degli Studi di Milano - Dipartimento di scienze veterinarie per la salute, la produzione animale e la sicurezza alimentare

Former Food Products (FFPs) represent a way by which convert losses from the food industry into ingredients for the feed industry, thereby keeping food losses in the food chain. However, as new ingredients, FFPs nutritional evaluation merits further investigations. In this respect, the aim of this study was to perform a nutritional evaluation of selected FFPs produced in the frame of IZS PLV 06/14RC PROJECT, in a FFPs processing plant. Six samples of mixed FFPs, all based on bakery products, have been analysed for dry matter (DM), ether extract (EE), Neutral Detergent Fiber (NDF), Crude Protein (CP) Crude Fiber (CF), starch and ash. Based on FFPs composition data, estimation of digestible energy (DE) and metabolizable energy (ME) values for pigs, have been calculated according to NRC (2012). ME/DE ratio has been also calculated. Mean nutrient contents were as follows: DM 920 (g/kg), EE 101 (g/kg DM), NDF 101 (g/kg DM), CP 100 (g/kg DM), CF 27 (g/kg DM), starch 524 (g/kg DM), ash 23 (g/kg DM). Estimated DE mean was 18.99 (MJ/kg DM), whereas estimated ME mean was 18.70 (MJ/kg DM). The ME/DE mean ratio was 0.98. Results obtained indicated that: i) FFPs considered in the present study exhibited a nutritional profile that is roughly equivalent to cereal grains; ii) fat content of tested FFPs was higher than conventional cereal grains, while starch concentration was comparable to conventional cereal grains; iii) FFPs tested were characterized by a valuable DE and ME density values for pigs, as also indicated by ME/DE ratio. However, DE and ME systems used may under/overestimate energy values due to the high lipid and starch content obtained in FFPs. Combining all obtained results, it can be suggested that FFPs may represent alternative high-energy ingredients for the feed industry.

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O6. New feed ingredients: the insect opportunity

RaamsdonkL.W.D.van,vanderFels-KlerxH.J.RIKILT Wageningen UR

In the framework of sustainability, animal health and/or feed safety, new ingredients for feed are desired and, to this end, a range of initiatives investigating such novel ingredients has been started. The opportunities range from herbs as replacements of growth promotors, second generation biofuel by-products, via refined products originating from grass, fresh

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water algae and marine weeds, to novel proteins, especially from insects. Within the EU, generally, a new feed ingredient should commit to legal constraints in terms of “yes, provided that”. In the case of animal proteins, however, a second constraint applies: “no, until”. These two principals will be presented with insect proteins as case.

A new feed ingredient is principally allowed when its safety commits to a range of legal limits for heavy metals, mycotoxins, pesticides, contaminants, microbiological safety etc. These constraints apply to products originating from insects as well. However, it should be considered that insects may behave differently from farm animals with regard to accumulation and/or excretion of contaminants. This has implications for the substrate given to insects, that are intended to be used as feed ingredients

Additionally, the use of animal proteins is subjected to the TSE legislation as well. Here, the principal is to reject the use unless an extra set of requirements is met.

The use of insect proteins is a good case to illustrate this balance between a positive, although restricted, modus, and a negative modus. In this complicated frame set, several opportunities can be explored for successful application. During the lecture the several aspects as applying to insects will be presented, the balance between the two modi will be explained and options for making this complicated case practicable will be discussed.


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Session 2: Traceability/authenticityoffeedmaterialsincludingAdditives

Keynote 2: Newsystemsforidentifyingpotentialrisksinthefeed chain

BreretonP.1,2, Miles T.1MojtahedV.11 Fera Science Ltd, York, UK 2 Institute of Agri-Food Research, University of Newcastle, UK

There is a long legacy of major food incidents being derived from animal feed (e.g. dioxins, BSE). Significant resources have been spent on addressing these risks and mitigating against repeat incidents. Less time has been spent on anticipating the nature and likelihood of future risks despite the huge savings in resources and reduction of brand damage that would result from the ability to target resources on where they are best needed. Anticipatory systems are required that can identify anomalies in data and metadata that might indicate atypical conditions in the feed chain. “Big data” systems are being developed that exploit perturbations in social media, the web, macro-economics and aim to provide stakeholders with warnings of possible future risks. In contrast, analytical testing regimes have usually focused on identification/monitoring of feed incidents and rarely provide warning that an incident is imminent or likely to happen. New developments in analytical technology and informatics now present the opportunity to incorporate food analysis systems within a risk mitigation strategy, identifying problems earlier in the incident pipeline.

Examples of “horizon scanning” systems will be presented together with an overview of how feed analysis methods can be developed to identify nascent feed risks.

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O7. Determinationofiodineandinorganicarsenicinfeed:analyticalmethods and risk assessment

Sloth J.J.1, Rasmussen R.R.1, Danier J.2

1 Technical University of Denmark, National Food Institute (DTU Food), Søborg, Denmark2 Bavarian State Ministry of the Environment and Consumer Protection, München, Germany

Iodine and inorganic arsenic are two important parameters in relation to evaluation of feed

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safety. Iodine is an essential element and both animals and human need adequate supply of this element to maintain a healthy life. However, excessive intake of iodine may also lead to adverse effects and therefore the exposure and consequently the content of iodine in feed should be monitored. Arsenic has a very complex chemistry with more than 100 different naturally occurring arsenic compounds identified – especially in samples from the marine environment. When it comes to feed (and food) safety evaluation, the emphasis is on the inorganic arsenic (iAs) fraction, since iAs is considered as the most toxic arsenic compound. The EU legislation has presently only maximum levels (MLs) for total arsenic in feed and several methods for the determination of total arsenic exist (incl standardized methods). However, arsenic speciation analysis is called upon to achieve specific data on inorganic arsenic in order to be able to perform a correct risk assessment. The recent developments in standardization of method(s) for inorganic arsenic determination will be an important tool for this task and will also open up for the possible introduction of specific MLs on inorganic arsenic in the feed legislation.

The present lecture will present the current status and latest development within methods of analysis for determination of iodine and inorganic arsenic in animal feed. Data from recent collaborative trials in regi of CEN TC327 WG4 working group on heavy metals, trace elements and minerals will be presented and discussed. Furthermore, future needs for further development of methods for iodine and arsenic (incl speciation analysis) in feed will be discussed and a few ideas on how to proceed will be elucidated.

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O8. OligonucleotidesofDNAasspecific,biologicaltracers

MollW.D.1,GruberA. 1, Brunner K.2

1 BIOMIN Research Center, Tulln, Austria, 2 University of Natural Resources and Life Sciences Vienna, Center for Analytical Chemistry, Department of Agrobiotechnology IFA-Tulln, Tulln, Austria

The objective of our work has been to develop a new technology for tracing and authentication of feed additives. The technology is based on double-stranded DNA oligonucleotides as tracer, and sequence-specific detection and quantification by real-time polymerase chain reaction (PCR) or digital droplet PCR.

DNA oligonucleotides with a length of 70 to 100 nucleotides were chemically ptimizatio, annealed to stable double-strands, and dried with carrier to make preparations of micrometer-sized particles with uniform loading of tracer DNA. Such preparations were used


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to trace feed additives, and subsequently feed, by dry mixing. For analysis, feed samples were extracted with aqueous buffer, extracts were purified, and the DNA oligonucleotide included as tracer was amplified and quantified by real-time PCR or digital droplet PCR.

After optimization of protocols, the use of microgram amounts of tracer DNA per ton of feed was sufficient for specific detection and quantification. This extremely low inclusion rate of tracer is enabled by the enormous amplification power of PCR: During analysis, the amount of tracer DNA is doubled by biological replication in each cycle of the PCR reaction, so that million-fold amplification and even detection of single molecules is possible. According to our results, the DNA tracer showed no loss of activity after feed pelleting or storage. Several features of tracing with DNA make the technology seem attractive: An unlimited number of tracers can be designed, produced and analysed with the same protocols, and mixed in any combination and ratio without influencing each other’s analysis. The tracer is exclusively composed of digestible biological macromolecules and used in such tiny concentrations that any effects on feed composition or quality are impossible. The DNA oligonucleotides are produced without use of recombinant DNA technology or fermentation, and therefore free of contaminations. We believe that implementation of our new tracer technology has potential to simplify and improve authentication and traceability of feed additives, which should be in the interest of the feed additive industry, control authorities, and animal health and welfare alike.

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O9. Theuseofspectroscopyinthefieldofmineralchelateanalysis.

CantwellC.,MurphyR.,ConnollyC.Alltech European Bioscience Centre, Sarney, Summerhill Road, Dunboyne, Co. Meath, Ireland.

ObjectivesTo develop and validate a solid state spectroscopic method for the quantitative analysis of mineral proteinates (Bioplex TM).

Materials and MethodsFourier Transform Infrared (FT-IR) spectroscopy was used to acquire spectra (1800-650 cm-1) from a range of specially prepared calibration standards and test samples. A multivariate analysis approach was required to extract useful information from the FT-IR spectra. Raw data were subjected to principal component analysis (PCA) to allow for the construction of appropriate scores plots and calibration curves. These were used in turn to predict the amount of bound mineral present in the test samples. A separate analysis of total mineral content by inductively coupled plasma optical emission spectroscopy (ICP-OES) allowed for the calculation of bound mineral as a % of total mineral content.

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Results and ConclusionsSpectroscopic analysis using FT-IR, together with a specific multivariate analysis of the data allowed for the accurate and precise measurement of bound mineral. This product-specific assay for mineral proteinates (Bioplex TM) was developed, validated and verified in accordance CRL-FA guidelines.

The non-destructive approach used overcomes some of the difficulties associated with assays that require dissolution or digestion of the test material before analysis. Certain challenges remain however, e.g. the product-specific nature of the assay requires specially prepared calibrants for individual products within difference organic trace mineral classifications. Although a similar FT-IR/PCA approach may be suitable for other products and preparations, validation and independent verification studies are required to ensure that it is “fit for purpose” and suitable for application under these conditions.

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O10. Traceabilityandauthenticityoffeedmaterialsusingnearinfraredspectroscopy and near infrared microspectroscopy: case studies

YangZ.,JiangX.,ShenG.,HanL.College of Engineering, China Agricultural University, Beijing, P.R. China

In recent years, feed safety related to human health is becoming an increased concern for consumers. NIRS technology plays an important role in feed industry for rapid determination of valuable constituents and parameters. This study discuss the potential of on-line NIRS and NIRS microscopy (NIRM) for traceability and authenticity of feed materials.

Case study 1: On-line NIRS can offer feed industry an effective and real-time analysis method. In this case, on-line NIRS was used to classify the geographical origins of fish meal. The results showed that the PLSDA model discriminant accuracy of calibration and validation were 100% and 89.74% respectively. Then competitive adaptive reweighted sampling (CARS) was used to select characteristic wavelength variables for PLSDA model, the discriminant accuracy for the validation increased to 94.87%.

Case study 2: The potential of NIRM to detect and identify meat and bone meal (MBM) in feed has been proved. But the accuracy is lower when MBM particles were overlapped by feed particles. A Markov random field based approach was developed using the iterated conditional mode (ICM) algorithm, integrating initial labeling-derived results from support vector machine discriminant analysis (SVMDA) and observation data derived from the results of principal component analysis (PCA). The results showed that MBM covered by feed could be successfully recognized with an overall accuracy of 86.59 % and a Kappa coefficient of 0.68.


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Case study 3: The identity and authenticity of feed materials are current topics in feed science. As driven by commercial interests, new illegal adulterants which are unknown to consumers and regulators emerge constantly. A non-targeted adulterant screening method based on NIRM is proposed. This study focused on the feasibility of non-targeted screening methods for the detection of adulteration in soybean meal. Six types of non-protein nitrogen were taken as examples. The results showed that the non-targeted screening method could screen out adulterations in soybean meal with satisfactory results.

The results obtained in these different case studies showed that NIRS/NIRM with chemometrics tools fits the traceability and authenticity of feed materials.

Acknowledgement:

This research was supported by the National Science and Technology Support Program (2014BAD08B11-2), Program of International S&T Cooperation (Project No. 2015DFG32170) and National Key Scientific Instrument and Equipment Development Project (2014YQ47037705). Some of samples used in this study were provided by the European Union’s Seventh Framework Program (FP7/2007-2013) under grant agreement 265702 (Quality and Safety of Feeds and Food for Europe, QSAFFE).

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O11. SpectroscopyappliedonfeedadditivesoftheEuropeanReferenceLaboratory: an important tool for traceability

SlowikowskiB.,OmarJ.,BoixA.,vonHolstC.European Commission, Directorate General Joint Research Centre, Geel, Belgium

Feed additives are an integral part of modern animal husbandry combining high performance in production with animal health and welfare. Such additives may not be put on the market without pre-market authorisation by the European Commission following a procedure established by Regulation (EU) No 1831/2003 [1]. When submitting a request for authorisation, the applicants need to send feed additive samples to the European Reference Laboratory for Feed Additives (EURL-FA). Currently, there are about 700 feed additives stored in the EURL-FA’s repository. Since the Regulations authorising these products often foresee corresponding identification criteria, there is strong need for analytical methods to enforce these provisions. On its own initiative, the EURL-FA subjected all feed additive samples to analysis by vibrational spectroscopic techniques. Near infrared (NIR) and Raman spectroscopy were applied because these methods have the unique potential of characterising the feed additives in a non-destructive manner, without the need of complicated sample preparation as already demonstrated in a previous publication [2]. The purpose of this study is to assess

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the capability of spectroscopy to establish whether specific authorisation criteria in terms of the characterisation of products are met. The study presented is based on the analysis of feed additive samples from different categories and functional groups. Five examples have been selected demonstrating the applicability of the vibrational spectroscopy to the intended purpose.

[1] Regulation (EC) No. 1831/2003 of the European Parliament and the council of 22 September 2003 on additives for use in animal nutrition. Off J Eur Union. Oct 18; L 268:29-43, lastly amended by Commission Regulation (EU) 2015/2294

[2] Omar J., Boix A., von Holst C. Differentiation of coccidiostats-containing feed additives by mid and near infrared microscopy. Food Additives & Contaminants: Part A, (2015) Vol. 32, No. 9, 1464–1474


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Session 3: Contaminants,undesirablesubstances,residues

Keynote lecture 3: EUPolicyonundesirablesubstancesinfeed: recent developments and outlook

Verstraete F. European Commission, DG for Health and Food Safety Brussels.

Directive 2002/32/EC of 7 May 2002 of the European Parliament and of the Council on undesirable substances in animal feed is the framework for the European Union action on undesirable substances in feed.

Following requests of the European Commission, the Panel on Contaminants in the Food Chain (CONTAM) from the European Food Safety Authority (EFSA) has completed in recent years several scientific opinions on contaminants in feed and food, reviewing the possible risks for animal and human health due to the presence of these substances in feed and food.

The outcome of these risk assessments has resulted in several changes to the EU legislation on contaminants in feed. A short overview of the recent changes with information on the considerations resulting in these legal provisions will be provided in the presentation.

Furthermore the presentation shall focus on following challenges for the future:

Increased prevalence of mycotoxins (and plant toxins) in feed.In recent years, an increased prevalence and a significant year-to-year variation of the presence of mycotoxins in feed and food in the European region can be observed. Climate change and extreme weather conditions are considered to be the main cause. The high levels of aflatoxin in the maize harvest 2012 and the high level of Fusarium toxins in the maize harvest 2013 and 2014 have resulted in problems for feed and food supply and safety. This situation entails specific challenges for farmers, feed and food manufacturers, traders and regulators to ensure the safety for animal and human health of feed and food while ensuring the supply of major staple fed and food such as cereals. This issue has been intensively discussed in recent years and the presentation shall provide more details on these discussions and explore some possible options on the way forward.

Acceptability criteria for detoxification of feed Commission Regulation (EU) 2015/786 of 19 May 2015 defines acceptability criteria for detoxification processes applied to products intended for animal feed as provided for in

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Directive 2002/32/EC of the European Parliament and of the Council. The Regulation applies as from 1 July 2017. This means that from 1 July 207 only detoxification processes which has been assessed and concluded by the European Food Safety Authority (EFSA) that the detoxification process complies with the acceptability criteria established in the Regulation, can be used for the detoxification of feed. Ttransitional measures are foreseen that for the detoxification processes for which the necessary information was provided to the Commission before 1 July 2016 but EFSA has not finalised the assessment by 1 July 2017,.

OutlookAn outlook shall be provided on the issues for which it is foreseen that these will be discussed in view of an update of the EU provisions on undesirable substances in feed following recent EFSA opinions or EFSA opinions to be expected the near future.

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O12. Levelsofundesirablesubstancesinmarinefeedingredients,commercialNorwegianfishfeedandfarmedAtlanticsalmoninthelast13-15years.

SandenM.,NøstbakkenO.J.,HemreG.I.,MaageA.,LunestadB.T.,EspeM.,LundebyeA.K.,AmlundH.,ØrnsrudR.National Institute of Nutrition and Seafood Research (NIFES), Bergen, Norway.

Fifteen years ago, Atlantic salmon diets were mainly composed of marine ingredients, while today the composition is a blend of marine and vegetable feed ingredients (~70% plant derived ingredients in Norwegian salmon feeds). This “blue-green” shift has reduced the level of several undesirable substances in the feed. The main objective of the present work was to evaluate levels of undesirable substances in marine feed ingredients, Norwegian fish feed and Norwegian farmed Atlantic salmon (Salmo salar L.). Through national surveillance programs more than 3400 fish feed samples were analyzed between 2003 and 2015 and more than 2300 samples of farmed salmon were analyzed between 1999 and 2011. The samples were analyzed for dioxins, PCBs, organochlorine pesticides (DDTs) and total arsenic.

Results on dioxins from the monitoring programs show that the concentration of dioxins (PCDD/F) and dioxin-like PCBs (DL-PCB) in fish oil show a general decline from 2003 to 2015, from a mean of 9.6 ng TEQ05 in 2003 to 4.7 ng TEQ05 in 2015. A decline in the concentration of PCDD/F and DL-PCB is also seen in salmon fish feed (n=606) from a mean of 2.8 ng TEQ05 in 2001 to 0.8 ng TEQ05 in 2015. The concomitant reduction in Norwegian farmed salmon (n=432) was from a mean of 2.0 ng TEQ05 in 1999 to 0.6 ng TEQ05 in 2011.


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During our period of analysis, the levels of dioxins, PCBs, DDTs and total arsenic in fish feed and Norwegian farmed salmon showed a general weak decrease. Our results will be presented and discussed in light of current upper maximum limits in feed and derived tolerable intakes. Finally, a brief overview of levels of new undesirable substances mainly derived from plant ingredients in Norwegian salmon fish feed will be presented.

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O13. Validationofaselectedpigreal-timePCRassayfortheDetectionofprocessedanimalproteinsinfeedingstuffs.

Fumière O.,MarienA.,MaljeanJ.,BerbenG.Walloon agricultural Research centre (CRA-W), Agricultural Product Valorisation Department/European Union Reference Laboratory for Animal Proteins in Feedingstuffs (EURL-AP), Gembloux, Belgium

The Commission Regulation No 51/2013 named the Polymerase Chain Reaction (PCR) as reference method of analysis besides the light microscopy for the determination of constituents of animal origin for the official control of feed. The validation of a PCR assay for the detection of ruminant DNA in feed (Fumière et al, 2012) already allowed the re-authorization of non-ruminant PAP in feed for fish since the 1st of June 2013 (Commission Regulation (EU) No 56/2013). Further steps to the lifting of the feed ban need the validation of additional PCR methods. The PCR method presented here was developed for the detection of pig DNA by the CRA-W. It was assessed and validated after its selection among six PCR methods put at the disposal of the EURL-AP.

The target is a multicopy one of 83 bp located in the mitochondrial DNA. The evaluation was conducted according guidelines established for validation of qualitative real-time PCR methods (Broeders et al, 2014) and international texts such as the Codex Alimentarius and ISO Guidelines (ISO/IEC 17025, ISO 24276:2006, ISO 21569:2005, ISO 21570:2005, ISO 5725) or other relevant publications in the field (Horwitz, 1995 ; von Holst & MacArthur, 2011). The method satisfies to the minimum performance criteria of specificity (tested against 45 animal and 7 plant species), sensitivity (fitness for the detection of minimum 0.1 % w/w of pig PAP in feed), efficiency (between 80 and 120 %), LOD (20 copies with a cut-off set at 5 copies) and robustness. As for the ruminant method, a transfer protocol using plasmid calibrants provided by the JRC-Geel must be applied to every PCR platform. Based on it, a cut-off value delimiting negative results from positive ones can be determined.

Finally, an interlaboratory study was designed to validate successfully the method. Following this validation study, a second interlaboratory study including the DNA extraction step

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was organized in the NRL network to check its correct implementation. Four NRLs did not provide results in due time but the results from the 24 remaining NRLs indicated an excellent performance confirming what was obtained during the validation study

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O14. Candidatestandardmethodforthedeterminationofcarotenoidsinanimalfeedingstuffs

SeranoF.,vonHolstC.,Vincent U.European Commission, Directorate General Joint Research Centre, Geel, Belgium

Carotenoids are feed additives classified in the category “sensory additives” and functional group “colorants” i.e. substances which, when fed to animals, add colour to food of animal origin (Annex I of Regulation (EC) No1831/2003). Legal limits of these compounds have been established in feed. Moreover, the authorisation of these compounds also includes corresponding Maximum Residue Limits (MRLs) in food matrices. Improved methods of analysis of carotenoids are therefore required in order to carry out official controls to ensure the verification of compliance with feed and food law, labelling claims, animal health and animal welfare rules (Regulation (EC) No 882/2004). Carotenoids present a variety of stereoisomers, due to the numerous conjugated double bonds and the cyclic end groups. The most important forms commonly found are geometrical isomers (E/Z); these are inter-convertible in solution and present different physical properties and in particular their absorption coefficient and the wavelength of maximum light absorbance (λmax) are different.

A multi-analyte HPLC method which enables the reliable quantification of authorised carotenoids in fish and poultry feeds and pre-mixtures has been successfully validated in-house. This new method has the important advantage that it is applicable to all the different sources of carotenoids (chemical and natural additives), which until now required the use of product specific methods. The new method involves a simple sample preparation, avoids the use of chlorinated solvents, is based on reverse phase HPLC and provides reliable quantification thanks to the isosbestic concept which makes it fit for the purpose of official control of carotenoids in fish feed. The method has been selected as candidate method for standardization under the third mandate, part III (M/523) of the European Commission to the European standardization body CEN.

References[1] Mitrowska K., Vincent U., von Holst C., J Chromatogr A. 2012 Apr 13; 1233:44-53.[2] Serano F., von Holst C., Vincent U., private communication, EURL-FA annual workshop 2015


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O15. Ananalyticalstrategyfortheearlyqualityandsafetyassuranceintheglobal feed chain

deJongJ.1,LópezP.1, Mol H.1, Baeten V.2,FernándezPiernaJ.A.2,VermeulenP.2, Vincent U.3, BoixA.3,vonHolstC.3,TomaniovaM.4,YangZ.5,HaugheyS.A.61 RIKILT-Wageningen University and Research Centre, Wageningen, The Netherlands2 Walloon Agricultural Research Centre, Gembloux, Belgium3 European Commission, Directorate General Joint Research Centre, Geel, Belgium4 Institute of Chemical Technology, Prague, Czech Republic5 College of Engineering, China Agricultural University, Beijing, China6 Institute for Global Food Security, Queen’s University Belfast, United Kingdom

ObjectivesIn the past decades, several major food safety crises, e.g. the dioxin crises in Belgium (1999), Ireland (2008) and Germany (2010) originated from problems with animal feed. Consequently, there is an urgent need for early detection of fraudulent adulteration and unintentional contamination of feed and feed materials. In the EU-funded research project QSAFFE, a strategy was developed that can be used by feed business operators and official inspectors to prevent dangerous goods entering the feed chain. This strategy was elaborated for two specific cases, viz. adulterations of (i) soybean meal with melamine and other types of adulterants/contaminants and (ii) vegetable oils with mineral oil, transformer oil or other oils.

Results and conclusionsThe first step of the strategy is screening at the feed mill or port of entry with non-destructive spectroscopic methods (NIRS and Raman). Afterwards, only for suspect samples it is necessary to perform post-screening and confirmation in the laboratory with MS-based methods. The spectroscopic screening techniques, that were developed in QSAFFE, are suitable for on-site and on-line applications. Portable equipment is becoming increasingly available. Currently NIRS and Raman are suited to detect fraudulent adulteration at relatively high levels but not to detect low level contamination. The potential application of the strategy for non-targeted analysis is demonstrated.High-resolution LC-MS and GC-MS methods, developed in QSAFFE, yield unambiguous confirmation and full-spectrum acquisition allows retrospective data analysis of suspect samples, adding extra value to the control strategy with respect to non-targeted approaches.

Acknowledgements The research leading to these results was conducted in the QSAFFE-project and has received funding from the European Union Seventh Framework Programme (FP7/2007-2013) under Grant Agreement no. 265702.

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O16. Naturalchloramphenicolincerealstraw–occurrence,transfertopigsandpossibleimplicationsinofficialcontrol

NordkvistE.National Veterinary Institute, Uppsala, Sweden

Detection of low levels of chloramphenicol in urine, muscle or organs from pigs might not be proof of illegal use of the drug. Naturally occurring chloramphenicol have been observed in cereal straw in North-western Europe. Studies of this and proof of concept for transfer to pigs have been conducted and possible implications for official control of food from animal origin will be discussed.

Based in the findings from the Netherlands and Sweden surveys were conducted where 105 samples of straw from the Netherlands, France, the UK, Germany and Denmark and 215 samples of straw from Sweden were analysed with respect to chloramphenicol. In 37 samples out of 105 and 54 samples out of 215 chloramphenicol was detected. The highest content was 32 µg/kg in a straw sample from Sweden.

The fact that pigs consume bedding straw possibly chloramphenicol naturally produced in the soil led to another experiment where growing pigs were exposed to chloramphenicol in low doses (4, 40 and 400 µg/kg) in order to examine if residues might be found in organs or urine. This experiment revealed a dose related increase of residues in muscle, plasma, kidney and urine, but no chloramphenicol was detected in liver (LOD <0.05 µg/kg).

This presentation is in essence based joint research efforts from RIKILT, Wageningen, Netherlands Food and Consumer Product Safety Authority, Utrecht, NL, the National Veterinary Institute (SVA) and the Swedish Food Administration, Uppsala, SE published in the following papers:

- Nordkvist, E.; Zuidema,T.; Herbes,R.G.;Berendsen,B.J.A.; Food Additives & Contaminants: Part A 2016, 33, 798-803. DOI: 10.1080/19440049.2016.1176450- Aspenström-Fagerlund, B., Nordkvist, E., Törnkvist, A., Wallgren, P., Hoogenboom, R., Berendsen, B. & Granelli, K. 2016 Distribution of chloramphenicol to tissues, plasma and urine in pigs after oral intake of low doses. Submitted for publication

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O17. ComputationaltoxicokineticmodelingforconsumerprotectionagainstcontaminantsinfeedandfoodattheFederalInstituteforRiskAssessment (BfR)

NumataJ.,MielkeH.,PirowR.,Müller-GrafCh.,SchafftH.,GreinerM.,Lahrssen-WiederholtM.Federal Institute for Risk Assessment (BfR)

Animal experiments on the transfer of undesirable substances along the feed and food chain deliver raw data, which strictly speaking only represent situations identical to the experiment. To be able to extrapolate from these specific data, a series of mathematical methods generally known as kinetics are required. The basic method for estimating kinetic parameters in animals requires simplified experiments with a single dose, assumes a one-compartment model and reads an elimination constant from a graphic of the decreasing (blood) concentration of a chemical species.

Computational toxicokinetic modeling takes the animal experimental results to create a generalized mathematical abstraction, so as to form a model of reality. Toxicokinetics models the absorption, distribution, metabolism and excretion of undesirable substances by means of mathematical equations. Computational toxicokinetics allows more freedom in the experimental design and choice of more physiologically realistic models. For the experimental design, it is possible to deal with variable doses, several washout periods and other deviations from the basic design. For the chosen model, it is possible to use several compartments and different kinds of saturable and non-saturable kinetic equations. The final toxicokinetic model allows predictions of the concentration of an undesirable substance in foods of animal origin from a known concentration in animal feed (or possibly also in the reverse direction).

Several in-house applications of toxicokinetic modeling for health risk assessment at the BfR will be presented:

- Using a priori models for planning feed to food transfer experiments: tetrahydrocannabinol (THC) in cow’s milk; bioavailability of lead in piglets; transfer of dioxins (PCDD/Fs) and dl-PCBs in cow’s milk.

- A posteriori evaluation of feed to food transfer experiments: several perfluoroalkyl acids (PFAAs) in edible tissues of pigs; in cow’s milk; and hen’s eggs.

- Environmental biomonitoring: Modeling PFAA-ubiquitary environmental concentrations using wild boar liver samples from Germany.

- Human biomonitoring: A toxicokinetic model for PFAAs for the monkey was adapted to human physiology. Given a fixed daily dose, this model allows predicting the blood concentration in steady state or after a specified dosing period.

- Migration of paper coatings: Kinetic behavior of diPAPs (a precursor of PFAAs) from human experiments and subsequent modeling.

- Dermal absorption of chemicals used in the textile industry: combining QSAR and human toxicokinetic methodology.

Some of these models have been developed in cooperation with RIKILT/Wageningen UR.

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Session 4: Theindustryperspective

Keynote lecture 4: FutureChallenges–FeedIndustryisabletohelpifPoliticsaccept

Radewahn,PeterGerman Feed Association / FEFAC; Bonn / Brussels; Germany / Belgium

1. Future challenges are of different kind. Not only the undispensable feed safety is to mention and to solve, but also questions of feed security and of food competi-tion have to be answered.

2. It seems to be crucial, to get an international agreement on how to deal with safety issues in feed. The “top-of-the-pyramide”-concept is key for feed- and food safety.

3. Feed industry has the tools and the power to face both challenges a. by using international free markets, b. by using newly developed components and materialsc. by reducing excremental emissions via reducing essential nutrients

without reducing performanced. by developing and using new feed additives, which enforce efficacy of

the used feed components

4. Third challenge to face is animal welfare and animal health, where animal nutri-tion has a key role to play

5. There is no clear borderline between obligations and opportunities for animal nutrition at the one hand and the opportunities of animal medicine. We need new sense giving discussions between the two sectors without professional com-petition, only facing the benefit of the sector

6. Rather free competition between some few politically set borderlines is the best way to find effective, intelligent and accepted solutions. It’s not the administra-tions duty to rule each and any relation between partners in the market.

7. There is a new style of discussing hazards and risks to be developed. We have to answer the question from a higher point of view, if each and any risk has to be avoided for health reasons? What do we want to accept as risks? This needs a social discussion from an overall point of view. We have to accept there is no zero risk!


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8. What are maximum limits for this and that contaminant worth? We should also accept that thresholds on each and any contaminant are only working if the risk manager decides to accept a certain level of risk. For that it is necessary for pol-itics as well as for administration, media and business to join forces together in communicating this remaining risk.

9. Analytical methods have to be developed or existing ones have to be improved to get the right results for the one or the other purpose. So even paper controls may be the method of choice. The method of control and the method of messurement has to be the right one, even in finding and assessing existing or emerging risks. Practical examples to be given.

10. If assessing the last minor risks leads us to situations where we get more and higher risks on another hand, public risk managers have to decide which way to go and to communicate this decision. Assessment of risk is not a self purpose and has to step back again behind the risk management decisions

11. The way forward is well described by the FEFAC-Vision 2030 that was published and introduced in July this year. It makes clear that sustainability is the way to face the challenges of future. It makes also clear, that animal nutrition is the spi-der in the net of sustainability.

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O18. SecureFeed:acollectivefeedsafetyapproachtomeetchangingdemands in the food chain

Gort, F. SecureFeed, Wageningen, The Netherlands

SecureFeed stands for the assurance of food safety of animal feed. Participants deliver their feed materials, compound feedingstuffs and feed additives directly to livestock farmers. SecureFeed develops and manages a joint system”, the “SecureFeed Assurance System” for monitoring and risk assessment of raw materials and their suppliers.

By sharing knowledge and information with partners in the food chain and external parties, SecureFeed is continuously working on strengthening the risk awareness of its participants and its risk approach. As an independent organization in the field of food safety of animal feed, it’s the point of contact for participants, chain partners and external parties.

The presentation will focus first on the elements of the SecureFeed Assurance System; products, suppliers and participants and how these three elements are closely linked.

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The next part of the presentation will focus on the results achieved in the first two years of SecureFeed’s existence.

In the third part the position of SecureFeed within the Dutch animal feed landscape will be discussed.

Finally, a short view into the future of this collective approach will be given, focusing both on the difficulties to be encountered but also the necessity of this approach.

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O19. UtilisingAnimalBy-Productsasasustainablefeedchoiceforfoodproducing animals

vanVuureC.Darling ingredients International, The Netherlands & EFPRA Belgium

EFPRA is Europe’s leading authority on the use, value and bio-security of edible animal fats and meat industry by-products. Its members work closely with regulators, livestock producers, meat processors and retailers across the EU to make best use of the 18 million metric tonnes of animal by-products produced every year.

The industry EFPRA represents recovers edible animal fats, valuable proteins and renewable energy at 450 sites, producing quality products for use in human food production, animal feeds, pet food and for the oleochemical, pharmaceutical, energy and construction industries.

When slaughtering an animal in Europe a significant amount is not classified as meat and / or not destined to be used in food. This is in practise about 30 – 40 % of the slaughtered animal, depending on the animal species,.

The rendering industry is always working to achieve the highest added value for all classes of slaughter by- and co-products to find the most optimal and sustainable value chain for the farmer, the slaughterhouse and the feed and food industry involved.

For sustainability reasons the feed industry, including compound feed for all food producing animals (pigs, poultry, aquafeed, etc.) together with the petfood and the fur animal industry, is the obvious choice for the use of animal proteins, fats and minerals. These ingredients provide the animals with highly digestible protein, fat and phosphorus. This is sustainable from the point of view of the use of European sources of those nutrients, but additionally, due to the high digestibility also positive for the environment by producing less manure.

Concluding: (Processed) Animal By-Products are a sustainable feed choice for food producing animals

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O20. Analysis of feed at farm and industry levels

BaetenV.,FernándezPiernaJ.A.,VermeulenPh.,VeysP.,DardenneP.CRA-W- Walloon agricultural Research centre (CRA-W),Gembloux, Belgium

The use of near-infrared (NIR) technologies for the detection of contaminants and undesirable substances in feed products is not widely practised. However, the papers published so far have demonstrated the unique advantages of the use of this fingerprinting technique in the continuing effort to give the stakeholders the means to detect contaminants at all stages of the feed chain. This is crucial in order to provide products that do not adversely affect animal and human health. For more than 15 years, the Walloon Agricultural Research Centre (CRA-W) has helped to demonstrate the potential of NIR spectroscopy in the detection, identification and quantification of animal, plant and chemical contaminants and undesirable substances.

This work has been based on several NIR technologies (i.e. classical NIR, NIR microscopy (NIRM), NIR hyperspectral imaging (NIR-HSI), on-line NIR and handheld NIR instruments). The achievements so far show that NIR, combined with adequate sample presentation and appropriate chemometric tools, has a specific place in strategies to ensure the analytical control of contaminants at the farmer, industry and laboratory levels. This has been done through the implementation of appropriate solutions in terms of limit of detection (LOD), robustness and sampling issues.

For instance, NIRM and NIR-HSI techniques have been used not only for detecting animal particles in compound feed, but also for developing methods for the full screening of particles in ground compound feed. They have been also proposed for detecting other types of plant and mineral contaminants. The NIR imaging system has been used successfully to detect, at the ppm level, the presence of ergot bodies and other plant contaminants and the botanical impurities in cereals. NIR HSI method enables a 250 g sample to be analysed in 1 min using the push-broom HSI system. The melamine crises also led to successful proposals to use NIR techniques (NIRS, NIRM and NIR HSI) to detect melamine in feed.

The potential of NIR technologies in the detection of contaminants is obvious. With instrument miniaturization and the reduction in instrument costs, it is clear that they will play an increasingly important role in ensuring safety in the food chain. The recent crises, linked to animal deaths and human health, demonstrated the need for a sensitive, reliable and rapid procedure for determining the presence of contaminants in agro-food products. The challenge, however, is to be able to detect the next crisis, whatever its sources.

The presentation will illustrates the last developments and achievements in the use of NIR techniques to detect contaminants in feed.

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O21. PMicrotracers (R) and Their Use in Coding Feed Products and Premixes inFinalFeedstoAssureFeedQualityandtoAssureAuthenticity.

EisenbergD.A.,EisenbergZ.S.Micro-Tracers,Inc. San Francisco, USA

Microtracers (R) were invented by Dr. Sylvan Eisenberg in 1955. Microtracers have been formulated as feed premix ingredients for more than 60 years in more than 700 milllion tonnes of feed with no adverse effect.

Microtracers “S” came first (colored uniformly sized salt particles), then Microtracers G (colored graphite particles), F (colored iron particles), RF (colored iron powder) and others. New SECURtracers are 100 micron particles, food grade, magnetically attractable, fluoresce, with 10 micron lettering on each particle. Some Microtracers are secret.

Microtracers are used widely to validate feed mixing and cross contamination control procedures and are included in GMP+ methods (Holland), ASAE methods (USA) and Tecaliman methods (France).

The Objective in their use as feed product ingredients is to allow immediate testing of feeds for the presence of the tracer as indicator for the coded active ingredient (vitamin, mineral, drug). This allows detection of manufacturing errors before feed is shipped. It is critical vitamin D3 be added to poultry feeds. This can be confirmed by testing feed for a Microtracer mixed with the vitamin. It also may be critical Nicarbazin not reach poultry breeder feed where it can be toxic or finisher feed where it can lead to tissue residues in poultry meat. This can be confirmed by testing every truckload of breeder and finisher feed.

A second Objective in formulating “Special” Microtracers in feed products is to code them and feeds containing them as proprietary. By testing feed samples, one can confirm the proprietary product was added to the feed.

Most Microtracers are iron based and magnetically retrievable from feed samples. Tests are completed in less than 2 minutes.

In 2015, feed manufacturers performed more than 500,000 tests detecting manufacturing errors that if not caught could have led to toxicity to animals, poultry or fish or to drug residues in finisher feeds that would have led to drug residues in foods.

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O22. FeedSafety:Regional,nationalandinternationalinitiatives.AChallengeforthefuture.

DejaegherY.BEMEFA-APFACA, Brussels, Belgium

Since the publication of the Food and Feed Safety Regulation in 2004 and 2005 we have seen a rapid development of private feed safety schemes influenced by several contaminations the feed sector was confronted with (dioxin crisis in Belgium, BSE-crisis in Germany, aflatoxin crisis in The Netherlands, ...). International organizations such as Fediol, Fefana, GAFTA discovered more opportunities for international feed safety schemes taking into considerations the international perspectives the companies are working in (oil crushers, chemical companies, ...). Next to these private initiatives several governments decided to (re)organize the public feed safety. For example UK, France, Belgium, Germany, Italy, … all decided to put a new structure in place , a Food Safety Agency, being in charge of food and feed safety. Depending of the member state a synergy (or no synergy at all) started to develop giving different results. The Directorate Health and Food Audits and Analysis (the former FVO: FOOD Veterinary Office) realized audits in several member states and came up with a bunch of remarkable recommendations. On top of that private stakeholders started also to develop initiatives with an immediate influence on the schemes: those initiatives, concerning feed safety but also quality or sustainability, are somehow interfering the existing models. How do companies have to position themselves in this complex world of schemes, methods of analysis, norms, monitoring plans? Is there a need for an international forum to improve mutual recognitions, to make exchanges possible? Or will Darwin succeed: the survival of the fittest?

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POSTERS SESSION 1

P1. Mediterraneankrillasanalternativeproteinsourceinfeed:traceelementsoccurrence.

SquadroneS.1,BrizioP.1,BattuelloM.2,NurraN.2, Mussat Sartor R.2, BenedettoA.1,PessaniD.2,AbeteM.C.1 Istituto Zooprofilattico IZSPLV, Torino, Italy

Krill are small crustaceans of the order Euphausiacea, found in all the world’s oceans. Krill has high protein content and omega-3 fatty acids, and constitutes a rich source of natural pigments, vitamins and other components highly valued in animal/fish feed. Although there are more than 80 different krill species around the globe, dried krill meals till now have been mainly produced from South Antarctic Krill (Euphausia superba). In Mediterranean Sea Meganyctiphanes norvegica is instead the more represented Euphausiacea species. The content of undesirable substances such as metals is closely related to the waters where krill is collected, and then our aim was to analyses trace elements contents in krill collected from Northwestern Mediterranean Sea in order to verify its suitability as an alternative safe source of proteins in feed. We monitored the concentration of 20 trace elements in krill samples; the sampling area was located offshore from the Italian coast, at the border between the Northern Tyrrhenian Sea and the Ligurian Sea. Krill samples were collected on September 2015. Stations was located on the continental shelf above bottom depth of 111 m. Determination of Al, As, Be, Cd, Ce, Co, Cr, Cu, Fe, Mn, Mo, La, Pb, Ni, Sb, Se, Sn, Tl, V and Zn was performed after wet digestion using acids and oxidants (HNO3 and H2O2). All metals were quantified by Inductively Coupled Plasma - Mass Spectrometry (ICP-MS Xseries II, Thermo Scientific, Bremen, Germany). Trace metals concentrations (mg Kg-1 wet weight) were the following: Zn (74,598) > Fe (27,756)> Al (27,658)> Cu (20,554)> Ni (1,667) > Pb (1,521) > Mn (1,275) > Se (0,689) > Cd (0,334)> As (0,300)> Cr (0,436) > Co (0,135)> V (0,097) >Mo (0,058) > Ce (0,038)> Sn (0,032) > La (0,016) > Sb (0,013). Be and Tl were < LOQ (0.010 mg Kg-1). NW Mediterranean krill samples have shown to be a good source of essential trace elements such as zinc, iron, copper, manganese and selenium. However the presence of the nonessential elements, in particular Al, Pb and Cd should be more deeply investigated, especially considering the high potential of zooplankton to be metals bio accumulator.


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P2. Response of Rainbow trout (Oncorhynchus mykiss) to peanut meal diets:effectsongrowthperformanceandsomehematologicalparameters

Acar Ü., TürkerA.Muğla Sıtkı Koçman University, Faculty of Fisheries, Department of Aquaculture, 48000-Mugla, Turkey

IntroductionThe price of fish meal (FM) keeps increasing because of the diminishing fish stock and increasing demand for the product. The increasing demand and progressive scarcity of FM in the international market boosted its price and launched the quest for reduction of FM in fish diets and the consequent search for alternative, acceptable protein sources. Peanut by-products, which remain after the extraction of peanut oil, can be used in fish feeds as an alternative protein source. The aim of the present study was to determine the effects of replacing FM with DPNM on growth and some hematological parameters of the Rainbow trout Oncorhynchus mykiss

Materials and MethodsIn a 60-day feeding trial, healthy cultured O. mykisss (mean weight ± SD = 71.69±1.21 g) were stocked 12 cages (1000–L) with 50 fish each. Defatted peanut pulp was obtained from Başpınar Fıstıkcılık, Osmaniye, Turkey. It was added to the feed at a rate of 0% (as control), 10%(DPNM10), 20% (DPNM20) and 30% (DPNM30) of fish meal.

Results and ConclusionsAt the end of the 8 week feeding period, the fishes that were fed with DPNM10 feed gained as much weight as those fed with the control feed prepared with fishmeal as the main source of energy. Feed conversion ratio (FCR) and specific growth rate (SGR) findings of rainbow trouts fed with trial feeds differed significantly from the control group as the peanut meal ratio used in feeds exceeded 10%. In the examination of the hematological parameters obtained from the rainbow trouts fed with feeds containing peanut meal at different ratios, it was detected that there were no significant differences compared to the control group. In conclusion,growth performance and hematological parameters results showed that peanut meal can be used as an alternative to fish meal up to 10% in rainbow trout diets.

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P3. Fungalcommunityassociatedwithlarvaeofblacksoldierflies(Hermetiaillucens) reared on vegetable waste

VarottoI.1, OttoboniM.2,SpranghersT.5,MartinE.1, Vallone L.2,MontagnaM.3,BandiC.4, Epis S.1, Eeckhout M.5,PinottiL.21 Università degli Studi di Milano - Dipartimento di medicina veterinaria, Milan, Italy2 Università degli Studi di Milano - Dipartimento di scienze veterinarie per la salute, la produzione animale e la sicurezza alimentare, Milano, Italy 3 Università degli Studi di Milano - Dipartimento di Scienze Agrarie e Ambientali Produzione, Territorio, Agroenergia, Milano, Italy4 Università degli Studi di Milano - Dipartimento di Bioscienze, Milano, Italy 5 Ghent University - Department of Applied Biosciences, Gent, Belgium

Insects are a promising part of future feed/food chains. Ongoing research on insects as feed is mainly focused on type of feed substrate to raise the insects (mini-livestock systems and substrates), nutritional values of the produced insects, diet formulation and the performances of animals, which have been fed with insects. However a key aspect of these materials is safety; in fact the use of insects or insect-derived flour incorporated in the feed may be a risk of potential zoonoses, pathogens, toxins and contamination with heavy metals. Accordingly, the aim of the present research was to study the intestinal fungal community of black soldier fly (Hermetia illucens) larvae raised on vegetable substrates, in order to characterize the presence of any toxin-producing fungi. The research has been focused on the isolation of yeasts and moulds from the intestinal content of larvae, fed on heat treated vegetable restaurant waste, and on the typing of isolates by ITS-RFLP and sequencing of the 26S rRNA D1/D2 domain. Results obtained indicated that the main toxin-producing fungi, according to literature, belonged to Pichia, Trichosporon, Rhodotorula and Meyerozyma genera. In a subsequent step of the experiment, the isolated yeasts were used for inhibition assays on sensitive yeast strains to test their killer phenotype, confirming the killer toxin activity of Trichosporon asahii yeast strain. Finally, the variability of the fungal community of each sample was also evaluated by metabarcoding using Next Generation Sequencing 454 technology of fungal ribosomal ITS region, confirming the influence of different growing substrates on the fungal community associated with the larvae of H. illucens.


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P4. CanGibelCarpmealuseinrainbowjuvenilesdietsasanalternativetoanchovy meal ?

Acar Ü ¹,KesbiçO.S.²,GültepeN.³¹ Muğla Sıtkı Koçman University, Faculty of Fisheries, Department of Aquaculture, 48000-Mugla, Turkey² Kastamonu University, Inebolu Vocational School, Sea and Port Management Program, 37500-Inebolu, Turkey³ Kastamonu University, Faculty of Engineering and Architecture, Department of Genetics and Bioengineering, 37200- Kastamonu, Turkey

IntroductionFish meal obtained from anchovy or herring used as protein source in commercial fish feeds, a situation that is both environmentally and economically unsustainable. Therefore, considerable research has been done investigating the use of alternative ingredients to replace fish meal in feeds formulated for carnivorous fish. Gibel carp one of the most abundant of the nonnative fish species in Europe. Recent studies show its very fast spread over the country and possible negative impacts on native fish communities. Therefore, this study have been conducted to determine the economic benefits of invasive species and potentials for aquaculture feed industryMaterial and methodsIn a 60-day feeding trial, healthy cultured Oncorhynchus mykiss (mean weight ± SD = 4.58±0.09 g) were stocked 9 aquarium (100 L) with 25 fish each. Gibelio meal was added to the feed partially replace fishmeal at a rate of 0%, 50% and 100%.Results and ConclusionsDietary gibelio meal replacement with anchovy meal has no negative effect on fish growth and some immune parameters. There were no significant different between all experimental groups compared with control diet. Also for some immune parameters of fish showed no significant differences among treatments such as serum lysozymee activity, myeloperoxidase activity and superoxidase dismutase activity. In conclusion, use of gibelio meal in diets of rainbow trout juveniles provides both advantages for more economic feedstuff in aquaculture industry and the fight against one of invasive species for European Countries.

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P5. Bioaccumulationofheavymetalsininsects:preliminarydata

Amato G.1, Ceppa L.1,GiovanniniT.1, Rizzi M.1,SquadroneS.1,AbeteM.C.1, Gasco L.2, MeneguzM.2, Marchis D.1

1 Institute Zooprofilattico of Piedmont, Liguria and Valle d’Aosta, Torino, Italy2 University of Turin, Department of Agriculture, Forestry and Food ( DISAFA ) Grugliasco, Italy

Objectives: Insect meal could be a source of proteins in feed. The aim of this project was to verify bioaccumulation of heavy metals in insects farmed on contaminated substrate.Material and Methods:Tenebrio Molitor larvae (10 days) were selected. Wheat bran was used as substrate larvae and analyzed by Atomic Absorption Spectroscopy (AAS).100 Tenebrio Molitor larve were divided in four groups and farmed on substrate contaminated with 6 mg/Kg (ppm) of Pb:Group A: larvae collected at half life cycle (20 days)Group B: larvae removed at half life cycle and placed on clean substrate for 6 days, then collected.Group C: larvae removed at half life cycle and placed on clean substrate until to the end of larva cycle (~40 days), then collected.Group D: larvae collected at the end of larva cycle.50 Tenebrio Molitor larvae divided in two groups were farmed on clean substrate (control groups):Group E: larvae collected at half life cycle.Group F: larvae collected at the end of larva cycle.All groups were analyzed by AAS to detect heavy metals (Pb, Be, Al, Co, Cd, Hg, Sn, Sb, Tl).Results and Conclusions:Wheat bran used as substrate showed a high level of Al (170 ppm) and a moderate level of Cd (0.43 ppm), whereas the Pb was absent. Significant level of Al (1.7 ppm) and a minimal level of Cd (0.053 ppm) were found in all groups, due to natural contamination of substrate. Pb levels were found in all groups of larvae grown on contaminated bran (Group A=0.48 ppm, Group B=0.030 ppm , Group C= 0.028 ppm, Group D=0.35 ppm). Six days of decontamination seems to determine a decrease in bioaccumulation of Pb. In fact the Pb level of Group C appear to be not significantly different from those of Group B. Pb was undetectable in control groups. Although Group D was breeded on contaminated substrate for a longer time, it showed a Pb level lower than Group A. Adult larvae appear to accumulate heavy metals less than young larvae, as it happens in shellfish.


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P6. Developmentandevaluationofreal-timePCRtargetsforthedetectionof insects in feed

GérardA.1,2, DebodeF. 1, Marien A. 1, Francis F. 2,BerbenG.11 Walloon Agricultural Research Center (CRA-W), Unit Traceability and Authentication,Gembloux, Belgium, 2 Functional and Evolutionary Entomology, University of Liège, Gembloux, Belgium

Insects are rich in proteins and could be an alternative source of proteins to feed animals. Numerous companies started the production of insects at small or larger scale as feed for chicken and fish. Most of the business models for feed production are based on the black soldier fly (Hermetia illucens) or the mealworm (Tenebrio molitor). In Europe, these novel feed are not yet authorized and products are commercialized outside Europe or eventually used as pet food (e.g. wild birds). For further authorization in Europe, many questions must be clarified concerning the presence of antinutritional compounds, the risk associated to pathogens, to residues (pesticides, antibiotics, heavy metals) and to allergens.

To authorize such products on the market, methods to detect if a product really contains insects and to authenticate insect products are also mandatory. European Commission Regulation No 51/2013 named the Polymerase Chain Reaction as a reference method to determine the constituents of animal origin in feed.

Targets focused on insects (target common to all insects) and targets specific to particular insect species are required. PCR methods are developed at CRA-W in this way. Among the methods developed, three of them already gave interesting results. The first one (81 bp) is specific to all insects excepting individuals from the Diptera order. The two others (94 bp and 114 bp) are specific to Tenebrio molitor. The specificity of the targets was tested against 45 insect species and on 10 commercial insect-based feed products (real-processed feed material samples). The sensitivity of the method was assessed through the AFNOR XP V03-020-2 standard approach using the LOD6 method. The three methods reached the recommended performance criteria (LOD≤ 20 copies).

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P7. Quantificationofruminantpapinfeed:whatcouldwedo?

vanDuijnG,Mensinga,JTriskelion BV, The Netherlands

As an ultimate result of TSE Road map 2, a new European Commission Regulation (Regulation EC 56/2013) came into force on 5 February 2013, which reforms the stringent rules on the use of processed animal proteins (PAPS) from non-ruminants (e.g. pigs and poultry) in feed. This Regulation amends the EU Regulation 999/2001 on transmissible spongiform encephalopathy (TSE Regulation) and is the product of the agreement reached between the Commission and technical experts from the EU Member States in July 2012.

The TSE Regulation was adopted in reaction to the poor control of meat and bone meal in the animal feed chain in the past. However, with the decreasing risk of TSEs throughout Europe it is recognized that there is now less need for these measures. The TSE Road map 2 was presented to the European Commission in July 2010. This road map reflects the final goal to at least partially lift the feed ban while assuring an extreme high level of food safety. The result being that, from 1 June 2013 onwards, PAPs from non-ruminants were deemed suitable for use specifically in farmed aqua feed only.

So far, the TSE Regulation imposed a general ban on the use of PAPs in the feeding of both ruminants and in non-ruminant animals, including fish and other aquaculture animals. Now, PAPs derived from non-ruminant animals can be used in aqua feed. The production of PAPs themselves is subject to the requirements as presented in the EU animal by-products legislation. In particular PAPs must only be derived from so-called ‘Category 3’ animal by-products (e.g. un-diseased carcasses and parts of slaughtered animals, including hides, skins, horns and feet), undergo pressure sterilization and be subject to controlled storage.

The relaxation of rules on PAPs in the feed chain was mainly based upon scientific opinions which found no TSE risk occurring from the provision of non-ruminant feed to non-ruminant animals, where ‘intra-species recycling’ (i.e. cannibalism) is avoided. Nevertheless, the European Parliament insisted that any move to revise the feed ban should be accompanied by specific methods in order to identify the species origin of proteins in animal feed containing PAPs so that intra-species recycling and the presence of ruminant PAPs can be excluded. These methods should also contribute to guarantee the minimization of the risk of cross-contamination during the production process performed under a regime of strict spatial segregation.

Several techniques based upon microscopy and spectroscopy have been considered for this purpose. Also, in order to allow a fast screening procedure on the PAP production- plants, rapid protein- specific tests based upon ELISA technology have been developed. By now,


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analytical procedures enabling the detection of DNA promise to be the most powerful. Within Triskelion BV various Real-Time PCR (Polymerase Chain Reaction) methods have been developed for the specific detection and discrimination of DNA derived from several species such as ruminants (inclusive cattle, goat and sheep), porcine and poultry. These methods even have been proven to be robust for analysis on PAP’s which, as a rule, experienced a treatment for 20 min. at 133o C and 3 bar. Further, in order to exclude the possibility of negative matrix effects, these methods were validated with a wide variety of samples including various types of meat- and bone meal from various sources. Also the applicability of the RT-PCR methods was shown for the traceability of different PAP’s in feed samples.

As an example, a RT-PCR method was developed for the amplification of a DNA sequence which is specific for the bovine alpha subunit precursor of the acetylcholine receptor. As this DNA target is highly abundant in ruminants (at least 200.000 copies per cell), the method is very sensitive. Further, as the DNA amplicon size is only 85 base pairs in length, extremely treated DNA material as present in PAPs will not lead to loss of respond in the RT-PCR experiments.

P8. Formerfoodproductssafetyevaluation:microbiologicalqualityandpackaging material residuals

TretolaM.,TirloniE.,BernardiC.,GirominiC.,OttoboniM.,CastricaM.,BaldiA,PinottiL.Università degli Studi di Milano - Dipartimento di scienze veterinarie per la salute, la produzione animale e la sicurezza alimentare, Milano, Italy

The use of alternative feed ingredients in farm animal’s diets can be an interesting choice from several standpoints, including safety. In this respect, the aim of this study was to investigate the safety features of selected former food products (FFP) intended for animal nutrition produced in the frame of IZS PLV 06/14 RC project by a FFP processing plant. Four samples of mixed FFP and 2 samples of raw FFP, all based on bakery products, were analysed for the enumeration of Total viable Count (TVC) (ISO 4833), Enterobacteriaceae (ISO 21528-1), Escherichia coli (ISO 16649-1), coagulase-positive Staphylococci (CPS) (ISO 6888), presumptive B. cereus and its spores (ISO 7932), sulphite reducing Clostridia (ISO 7937), Yeasts and Moulds (ISO 21527-1) and the presence in 25 g of Salmonella spp. (ISO 6579). On the 2 raw FFP (one mashed and one pelleted) presence of undesired ingredients which can be identified as remnants of packaging materials has been evaluated by microscopy according to Raamsdonk et al. (2012) and Marchis et al., (2016). Mean TVC and Enterobacteriaceae were 4.92 and 3.6 Log CFU/g, respectively. levels of B. cereus and its spores were below 3 Log CFU/g. Yeasts and Moulds were detected in all the samples, with counts around 4 and 3 Log CFU/g, respectively. Clostridia were found in very low counts in few samples, while E. coli and CPS were always below the detection limit (2 Log CFU/g). Among the samples

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collected Salmonella was always absent. When remnants of packaging materials has been considered, the inclusion was 0.005% (w/w) in the mash former feed and 0.02% (w/w) in the pelleted one. Of note, in both samples the packaging remnants were observed mainly from the 1-millimeter sieve mesh fractions. Based on this results it can be concluded that FFP are wholesome, even though some improvements in FFP processing, at the feeding plant, can be useful in further reducing their microbial loads and impurity.

P9. Influenceofapplepomaceinclusionontheprocessofanimalfeedpelleting

VukmirovićĐ. 1,MaslovarićM.2,PezoL.3,ČolovićR.11 University of Novi Sad,Institute of Food Technology, Novi Sad, Serbia2 Institute for Science Application in Agriculture, Belgrade, Serbia3 University of Belgrade, Institute of General and Physical Chemistry

Apple pomace (AP) is the main by-product of apple juice production. Fresh, ensiled or dehydrated AP is well accepted by the animals and this by-product has a large potential to be used as a feed-stuff. It has low protein and fat content but could be a good source of soluble carbohydrates, dietary fibers, minerals and polyphenols. Dried AP could be ground and used in the industrial production of animal feed. According to existing research data, 10% of AP could be included in rations for broilers, up to 20% in rations for fattening pigs, and more than 30% in dairy cows’ rations. Predominant principle in modern animal breading is to use animal feed in pelleted form, especially in poultry and pig breeding. The aim of this research was to study the effect of AP inclusion on pelleting process and physical quality of produced pellets. For this purpose three model mixtures were prepared composed of corn and sunflower meal as conventional feed-stuffs, and AP as an alternative feed-stuff. The AP was included in model mixtures at three inclusion levels (0, 10, and 20%), and pelleted using three thicknesses of the pellet press die (18, 24, and 30 mm), and three moisture contents of the material (13, 15, and 17%). Addition of 10 and 20% of AP to the model mixtures increased energy consumption of pellet press by 17.5 and 26.7%, respectively, compared to the mixture without AP inclusion, but pellet quality (expressed as pellet durability index) was increased by 40 and 50%, respectively. Using the thinnest die (18 mm) resulted in poor quality of pellets produced without AP, while inclusion of AP in the mixture resulted in good quality of the pellets produced using the thinnest die. This suggests that AP, besides nutritive value, has very good binding properties. Even though AP inclusion increases energy consumption of pellet press, at the same time it enables obtaining good quality pellets by using thinner die which significantly decreases energy consumption of the pellet press.


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P10. Bioconversionofbrownseaweedbyblacksoldierflylarvae–effectsonnutritionalcompositionandfeedsafetyconsiderations

Lock, EJ. 1,Biancarosa,I. 1,2,Liland,N.S.1,Belghit,I.1,Amlund,H.3,Ørnsrud,R.3, Bruckner, C4,Araujo,P1,WaagbøR. 1,2

1Department of Requirements & Welfare, National Institute of Nutrition and Seafood Research, Bergen, Norway2University of Bergen, Department of Biology, Bergen, Norway3Department of Risk and Benefit Fish, National Institute of Nutrition and Seafood Research, Bergen, Norway4Norwegian Institute of Bioeconomy Research (NIBIO), Bodø, Norway

Introduction:Black soldier fly (BSF, Hermetia illucens) larvae are efficient converters of carbohydrate-rich plant material into protein and lipids, suited for feed and food applications. Micronutrients as well as contaminants ¬can readily accumulate in BSF larvae. As such, the feeding medium has a direct effect on the nutritional value as well as the safety aspects of the insect products. Whilst insect products have a great potential from a nutritional standpoint, investigative studies on the transfer of contaminants from media to BSF larvae are scant. Seaweed is an underused resource that contains valuable nutrients like eicosapentaenoic acid (EPA) and iodine, commonly associated with marine ingredients. However, seaweed can also contain considerable amounts of minerals, heavy metals and (inorganic) arsenic.

Objective: The objective of this study is to investigate the possible transfer of nutrients, heavy metals and (inorganic) arsenic from the brown alga (BA) Ascophyllum nodosum to BSF larvae.

Materials & Methods:A growth trial with BSF larvae was performed with a regression design, where 10 to 100 % of a plant-based growth media was replaced with ground seaweed, the brown alga Ascophyllum nodosum.

Results and Conclusions:EPA, iodine and vitamin E concentrations increased in whole insects when dietary BA inclusion increased. Simultaneously, arsenic and cadmium were transferred from seaweed to the whole insects. A broader understanding of the effect of the feeding media on the larvae composition can help to tailor BSF larvae into a nutrient profile suited for specific feed or food applications, without compromising the safety of the insect products.

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P11. CharacterisationofpolydispersenanoTiO2byfield-flowfractionation

JoneOmar,ChristophvonHolstandAnaBoixEuropean Commission, Directorate General Joint Research Centre, Geel, Belgium

The European Commission (EC) published in 2011 a Recommendation on the definition of nanomaterials to facilitate determination when a material should be considered as nanomaterial for regulatory purposes in the European Union. According to this Recommendation, nanomaterial means ‘a natural, incidental or manufactured material containing particles, in an unbound state or as an aggregate or as an agglomerate and where, for 50 % or more of the particles in the number size distribution, one or more external dimensions is in the size range 1 – 100 nm’[1]. The proper implementation of this definition triggered the need of developing and validating characterisation and quantification methods for nanomaterials.

Titanium dioxide is one of the most widely used nanomaterials which can appear in a broad variety of final products such as paint pigments, toothpastes, sunscreens, coffee creamers, food colorants, etc. due to its valuable properties. Field-Flow Fractionation (FFF) is a very useful technique for the characterisation of such polydisperse materials due to its powerful size-based separation capacity; moreover, when coupled to the relevant detectors it allows for a thorough characterisation of the size of the nanoparticles. In this study an approach based on the use of experimental design for the optimisation of an FFF method for the characterisation of polydisperse TiO2 is presented. The work included the optimisation of the required dispersion protocol of the test samples and of the FFF conditions. The optimised method is applied for the characterisation of twelve commercially available TiO2. Finally, since the implantation of the EC Recommendation requires expressing the particle size distribution (PSD) in terms of number of particles per size class compared to the total number of particle in the material (e.i. number-based PSD), an approach for estimating the number-based PSD of the studied materials is presented.

[1] European Commission, Commission Recommendation of 18 October 2011 on the definition of nanomaterial, Official Journal of the European Union. 2011/696/EU, (2011) 38-40.


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POSTERS SESSION 2

P12. Nearinfraredspectrometryofdiagnosticallysignificantfattyacidsinfood feed and serum

Kalinin A.V. 1,KrasheninnikovV.N.1,SviridovA.P.2,TitovV.N.31 Institute for Spectroscopy, RAS, Moscow, Troitsk, Russia2 Institute of Photon Technologies, Research Center of Crystallography and Photonics, RAS, Moscow, Russia3 Russian Cardiology Research and Production Complex, Health Ministry, Moscow,Russia

The control of the content of fatty acids (FA) (primarily palmitic saturated, oleic monounsaturated and the sum of polyene FA) in such media as food, feed and blood is socially and clinically significant. We developed inexpensive, high sensitive and reliable optical method of quantitative determination of the content of mentioned substances in natural media based on the use of portable diffraction grating spectrometer with linear InGaAs array sensor and its confirmationwith IR Fourier transform and Raman spectra. The evidence of selectivity and reliability of FA quantitative analysis was obtained by the calibration (regression) on the projections to latent structures using standard mixtures of oils and fats. The wavelength working region of 1.1 -1.9 micron used for a portable diffraction spectrometer was proved by comparing with different calibration models for FTIR spectrometer in parts of the near infrared spectrum (for various overtones and combinations of FA absorption). Multiple correlation coefficient values of 0.89, 0.85 and 0.96 and standard error of about 0.5%, 1.4% and 0.4% for palmitic, oleic and the sum of polyene FA, respectively, were obtained. The ability to determine the content of mentioned FA in butter, spead, veterenary supplements and serum was confirmed

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P13. Effectsofpomegranteseedoilongrowthperformanceinrainbowtrout(Oncorhynchus mykiss)

ÖzdemirR.C.1,KesbiçO.S.2 , Acar Ü.3,GültepeN.4, Fazio F.5

1 Kastamonu University, Fisheries Faculty, Department of Aquaculture, Kastamonu2 Kastamonu University, Inebolu Vocational School, Sea and Port Management Program, Inebolu, Turkey3 Muğla Sıtkı Koçman University, Faculty of Fisheries, Department of Aquaculture,Mugla, Turkey4 Kastamonu University, Faculty of Engineering and Architecture, Department of Genetics and Bioengineering, Kastamonu, Turkey 5 Dipartimento di Scienze Sperimentali e Biotecnologie Applicate – Laboratorio di Cronofisiologia Veterinaria e Complessità biologica, Facoltà di Medicina Veterinaria, Università di Messina, Messina, Italy

Introduction The use of antibiotic as immunostimulants can be harmful to animals, consumer and enviroment. Therefore, the researchers have been focused on natural products to replace the antibiotics in fish feeds. Pomegranate seed oil (PSO) may serve as an alternative feed additive for growth promotion in fish. Therefore the present study was aimed to examine the effects of different levels of Pomegranate seed oil as a feed additive on growth performance of rainbow trout.

Material and Methods The rainbow trout (6.79±0.07 g) were placed in 12 aquariums and the aquariums volumes are 100L. The mean initial stocking density was 20 fish per aquarium. PSO was added dosage of 5, 10 and 20 g kg -1 in commercial trout feed. Fish were fed two times a day ad libitum for 60 days.

Results and Conclusion All experimental diets were accepted by the fish. The feed supplemented with PSO have impact on fish growth performance. The experimental groups which include 5 and 10 g kg-1 PSO showed lower feed conversion ratio value compared with control and 20 g kg-1 PSO group. Relatively growth rate (%) rate is positively effected by 5 and 10 g kg-1 groups compared with control and 20 g kg-1 PSO groups. As a result PSO supplementation in the diets especially 5 and 10 g kg-1 improve growth performance in rainbow trout.


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P14. Proteomicsapproachesforspeciesandtissuesspecificdifferentiationofprocessed animal proteins in aquafeeds

RasingerJ.D. 1,MarbaixH.2, Dieu M. 2, 3, Fumière O. 4, Mauro, S. 5,PalmbladM.6, Raes M. 2, Berntssen M.H.G. 1

1 National Institute of Nutrition and Seafood Research, Bergen, Norway2 URBC-NARILIS, University of Namur, Namur, Belgium3 MaSUN, Mass spectrometry facility, University of Namur, Namur, Belgium4 CRAW, Valorisation of Agricultural Products Department, Gembloux, Belgium5 CRA-W, Biotechnology Department, Gembloux, Belgium6 Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, the Netherlands

Non-ruminant processed animal proteins (PAP) are again allowed to be used in aquafeeds. Currently, light microscopy methods are used to detect PAP when the feed is not supposed to contain PAP or blood products. Conversely, a European Union Reference Laboratory (EURL) validated polymerase chain reaction (PCR) based method is used for ruminant DNA-detection when the feed is known to contain PAP or blood products, as indicated from the declaration or the labelling. However, PCR does not allow for a differentiation of cellular origin, and hence cannot be used for tissue specific (i.e. bone, feather etc.) identification. Potential need for identification of sources (tissue) of contamination have sparked the development of analytical methods complementary to PCR. Protein-based methods are already well established, and the latest developments in the field of proteomics and proteomics bioinformatics have strongly facilitated the use of these tools in the study of food safety and food authenticity. We investigated different proteomics tools for the detection of species and tissue specific peptide marker candidates of prohibited PAP material of bovine and ovine origin, and allowed PAP material such as porcine (blood, bone-carcass, greaves) and poultry (blood, bone-carcass, feather) origin. We found that irrespective of sample preparation, gel-based proteomics tools are inappropriate when working with PAP. Gel free shotgun proteomics approaches, on the other hand, are able to provide quality data that can successfully be mined for species specific peptide markers using peptide mass fingerprinting and machine learning. Yet, this approach still fell short when tissue specificity was the goal. Satisfying results in terms of both PAP species and tissue specificity were achieved only when gel-free shotgun proteomics in combination with spectral library based data analysis methods were used. Current work is focused on the standardization of the protein extraction and digestion procedures for regulatory use and the creation of an extensive freely available PAP spectral library reference collection.

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P15. CarotenoidprofilingastoolforverifyingeggproductionclaimsintheUK?

Heinrich K.,PyeC.,DonarskiJFera Science Ltd, Food Quality and Safety, York, UK.

Objectives:To develop and validate a method for the determination of production origin for eggs by (i) developing a carotenoid profiling method for hen eggs (and feed) and (ii) analysing hen eggs and feed from different productions methods (caged, barn, free range and organic) and regions in the UK (England, Northern Ireland, Scotland and Wales).

Materials and Methods:As part of the Defra project, FA0159, 18 eggs and a feed sample were collected by poultry officers from each of 16 different UK farms. On arrival at Fera Science Ltd, the egg samples were stored at 3 conditions (+4, +19 and +23 °C) for a 5 week period, generating 6 time points (week 0, 1, 2, 3, 4, 5).Individual eggs were taken from each farming system for week 0, 3 and 5, when stored at +4°C. In addition, at week 5, eggs stored at +19°C and +23°C were also sampled in order to give an indication of whether carotenoid profiles are affected by elevated temperatures. A total of 78 egg samples and 14 feed samples were analysed for carotenoids (capsorubin, capsanthin, lutein, zeaxanthin, citranaxanthin, β-cryptoxanthin, ethyl-8’-apo-β-carotene-8’-oate, β-apo-8’-carotenal, β-carotene and canthaxanthin) by HPLC-UV/VIS.

Results and Conclusions:Capsorubin and canthaxanthin were not detected in any of the egg samples, whereas capsanthin, lutein/zeaxanthin and β-carotene were detected in all them; citranaxanthin and β-8’-apo-carotenal were absent in organic ones. Only lutein/zeaxanthin were present (≤ 1 mg kg-1) in all feed samples. This suggests that the carotenoids are accumulated within the egg yolk, as the concentrations ranged from 0.8 to 12.7 mg kg-1 for week 0 samples. The absence of citranaxanthin and β-8’-apo-carotenal in organic feed samples was also confirmed.Differentiation of organic farming practice using the procedure reported by van Ruth et al (2011) was not possible due to lack of the canthaxanthin marker in UK feed.But the presence of citranaxanthin and β-8’-apo-carotenal in non-organic egg samples might be used, as a marker in UK eggs, to differentiate them from organic ones.


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P16. Verificationofthegeographicaloriginofgrainmaizeusingnon-targetedFourier-transforminfraredspectroscopyspectroscopy

RiedlJ. 1, Achten E. 1, Schuetz D. 2, Fischer M. 2,Lahrssen-WiederholtM.1, Fauhl-Hassek C. 1 1 Federal Institute for Risk Assessment, Berlin Germany2 University of Hamburg, Hamburg School of Food Science, Institute of Food Chemistry, Hamburg, Germany

The verification of geographical origin of food and feed materials becomes more and more of interest to support the integrity of food and feed products. In particular, in ban situations, it has been observed that feed material of a suspicious area is subjected to misdeclaration to avoid these strict rules. In the feed sector, typically record-based traceability systems are used to inspect the authenticity of a product, but analytical approaches for geographical origin verification are still rarely in place. However, first research projects on analytical authentication of feed materials have shown promising results, e.g. the QSAFFE project (www.qsaffe.eu). Moreover, in several research projects, the advantages of non-targeted approaches for authentication purposes have been demonstrated. Hereby, a non-targeted approach means that the whole spectral information is used as a characteristic fingerprint of the sample and multivariate statistics are applied for the interpretation of these fingerprints. In the presented study, a non-targeted screening approach is applied to verify the geographical origin of grain maize. Grain maize is an important, globally marketed feed material, which is also a raw product for several process chains. Therefore, it is a suitable demonstrator for authenticity testing on different spatial scales, namely within Germany, Europe and around the world. Samples from different regions of the world were investigated using fast attenuated total reflection Fourier-transform infrared spectroscopy (FT-IR) followed by multivariate statistics to build classification models for the verification of the geographical origin of unknown samples. Classification results of two types of samples preparation will be compared, the ground maize measured directly and the extracted oil fraction. In future other spectroscopic and spectrometric methods will accompany FTIR spectroscopy for complementary data acquisition and data fusion.

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P17. Homogeneitycharacterizationoffeedpowderusinghyperspectralimaging and chemometrics

FernándezPiernaJ.A. 1, Vincke D. 1, Ouizem Z. 2,DardenneP.1, Baeten V. 1

1 Food and Feed Quality Unit, Valorisation of Agricultural Products Department, Walloon Agricultural Research Center, Henseval Building, Gembloux, Belgium.2 Gembloux Agro-Bio Tech, University of Liège, Gembloux, Belgium

Objectives-In many industrial fields such as pharmaceuticals, process engineering and food/feed processing, handling and mixing of powders is an important step where many problems can happen, especially when the objective is to choose the right working parameters to perform a correct homogenization. In the case of the food and feed industries, the characterization of the homogeneity of mixtures of powders by the use of a rapid and sensitive method is a priority. Several studies have been published, which are mainly linked to the procedure to obtain mixtures of powders and the problems of non-uniformity that can be faced at industrial level.In this work, the aim is to propose a fast and easy methodology to define and characterize the homogeneity of feed mixture powders. This is an important requirement of the European Reference Laboratory for Animal Proteins (EURL-AP) when preparing inter-laboratory studies where homogeneity of samples should be assured. Here to characterize such homogeneity of samples, the proposed procedure includes the use of a NIR hyperspectral imaging system combined with an image treatment PCA-based protocol [1] as an alternative to well-known statistical methods, such as auto-correlation functions or variances. Material and Methods-Near infrared hyperspectral imaging combined with chemometrics has the advantages of its speed, robustness, and especially allows analyzing the products without major operations preparations. In one analytical step, it is possible to get spatial and spectral information of the mixture.Results and Conclusions-This study has allowed us to characterize the homogeneity of binary mixtures of feed ingredients as well as check the influence of homogenization duration. The results obtained indicate that degree of homogeneity of a feed powder mixture is influenced by several factors namely the homogenization time, the physical and chemical characteristics of the particles and the proportions of components. Moreover, the results have been modified in order to take into account the recommendations done by the working group for blend uniformity testing [2]. The results have been compared to other techniques to determine homogeneity based on PCR, classical microscopy and NIR microscopy.This work was partially performed within the framework of the European Reference Laboratory for Animal Proteins (EURL-AP).[1] H. Berthiaux, V. Mosorov, , L. Tomczak, C. Gatumel, J.F. Demeyre, Chemical Engineering and Processing, 45, 397 (2005);[2] Trudy, J. of GXP Compliance, 12 (1), 46 (2007).


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P18. Microscopicexaminationofteleostfishdiversity

Van RaamsdonkL.W.D.,VliegeJ.J.M.,RheeN.vande,PinckaersV.G.Z.RIKILT Wageningen UR, The Netherlands

Fish meal is a regular ingredient in animal feed. Besides the prohibition in feed for non-weaning animals, it is generally allowed. Notwithstanding this situation, further attention for the proper detection and examination in the framework of the ban on animal proteins is required, for several reasons:

• in specific cases it appears to be difficult to discriminate between material from fish and from terrestrial animals. Especially bone fragments from salmon can mimic those of terrestrial animals.

• in certain occasions detection at the required level of 0.1% seems to be difficult. This is indicated by recent results of proficiency tests.

• the species-to-species ban does apply to off all resulting from hatcheries or fish farms. This by product is not allowed to be fed to fish of the same species.

For all those cases it is advantageous to have more insight in the diversity of fish meal particles, especially those originating from the skeleton, since bone fragments are the primary targets of microscopic examinations.A set of 19 species originating from seven teleost orders was investigated. Specimens were divided in parts, i.e. skin, head, fins and body were separated and dried. Meat was largely removed from the body part before drying. Gills and scales were extracted from the head and skin, respectively. Bone fragments, gills and scales were investigated.A major aspect for the recognition of fish bone fragments is the situation that fish of more advanced orders generally show an acellular bone structure. When present in a sediment (fraction of a compound feed with high specific density, i.e. minerals and bone fragments), it might be difficult to discriminate between certain minerals and acellular bone fragments.The research resulted in an overview of scale shape and texture, gill structure, and typification of the diversity in bone fragments per species. The results are supportive in further addressing the proper detection of fish meal, as a whole or for species identification, and for discrimination with other types of materials in compound feeds.

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POSTERS SESSION 3

P19. Truenon-targetedanalysisofantimicrobialactivecompoundsusingBio-assaydirectedscreeningandliquidchromatographyhighresolutionmass spectrometry

WeghR.S.,StolkerA.A.M.,ZuidemaT.,BerendsenB.J.A.,DriessenW.D.M.,PikkemaatM.G. RIKILT, Wageningen University and Research Centre, Wageningen, The Netherlands

Microbial growth inhibition tests are widely used as a screening approach for the detection of antibiotics in animal feed. Samples, which show inhibition in the microbial test, are measured with group specific LC-MS/MS methods for confirmation. However, in some cases the active compound cannot be identified. In this study an alternative approach is developed to identify possible unknown active compounds in feed samples. The approach is based on four steps 1) sample extraction, 2) fractionation of the sample extract under different conditions, 3) identify which fractions shows microbial growth inhibition followed by step 4) LC-HRMS analyses to identify the unknown compound by data reduction and using accurate mass database searching. In order to detect unknown compounds with antimicrobial properties with a wide variety of physical properties, a generic sample preparation method is necessary and thus only sample extraction is applied, without any clean-up. Hugh data reduction is obtained using different fractionation conditions which introduce matrix variation, resulting the possibility to discriminate between relevant and non-relevant data. The coupling with databases increases the possibility to identify the compound of interest. The presented method has been tested with several interesting applications resulting corrected identification and confirmation of the unknown antimicrobial active compound.


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P20. Developmentandvalidationofamethodfordetectionandquantificationofpackagingresiduesinfeedfromformerfood.

Marchis D. 1,GiovanniniT.1, Amato G. 1, Ceppa L. 1,PinottiL.2, Gili M. 1

1 Institute Zooprofilattico of Piedmont, Liguria and Valle d’Aosta, Turin, Italy 2 University of Milan, Department of Health, Animal Science and Food Safety, VESPA, Milan, Italy

Objectives: Former food products are used to produce commercial complementary feed. A gravimetric method for quantification of packaging residues in feed from former food was developed and validated according to Regulation (EC) 882/2004. Materials and methods: 100±1 g of material was weighted, then it was sieved at mesh sizes of 2 mm, 1.4 mm, 1.25 mm, 0.5 mm. Every fraction was examined in a large plate with a large magnifying glass, and all particles of packaging residues were picked up with a pair of tweezers. Particles were weighted, then placed in a 250 mL beaker and defatted with 100 mL tetrachloroethylene (TCE) for 5 minutes. TCE was decanted and the remains allowed to dry under a hood for at least 2 hours. Residues were finally weighted and percentage reported (w/w). Validation criteria were examined using protocols set out in European legislation: specificity, accuracy, precision, repeatability, reproducibility, limit of quantification (LOQ). Specificity was evaluated on 20 blank feed samples. 0.05% w/w was the assigned value for LOQ. Precision was evaluated as intralaboratory reproducibility: 3 replicate measurements of 3 different operators on 6 samples spiked at LOQ level (0.05% w/w packaging residues -plastics, paper, aluminium); Accuracy was calculated on the precision 18 samples. Results and ConclusionA gravimetric method was developed, to detect and quantify packaging residues in feed. Data reported showed that it is suitable for routine analysis of packaging residues in feed with good recovery and specificity and repeatability results. Validation results: mean recovery 90%, repeatability RSD 6.26%, reproducibility RSD 6.4%, accuracy SD 5.71%.A control chart was elaborated using validation data that can be routinely used to check the maintenance of repeatability and accuracy conditions. This methods fits for purpose, both for official controls in laboratories, and for in house monitoring processes in former food industries.

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P21. Multi-mycotoxinsemi-quantitativescreeningofanimalfeedsamplesthrough the use of biochip arrays

PlotanM.,DevlinR.,PorterJ.,BenchikhM.E.,SibandaL.,RodríguezM.L.,McConnellR.I.,FitzGeraldS.P.Randox Food Diagnostics, Crumlin, United Kingdom.

Introduction. The main known groups of mycotoxins are aflatoxins, fumonisins, ochratoxins, trichothecenes A (T-2 toxin, HT-2 toxin), trichothecenes B (deoxynivalenol), and zearalenones. Co-occurrence of mycotoxins in a wide range of samples has been previously reported globally. For food and feed safety the determination of these mycotoxins is extremely important. Biochip Array Technology (BAT) allows the simultaneous semi-quantitative determination of a broad range of mycotoxins from a single sample to accommodate the lowest established global guidance limits for a particular feed type. BAT enables the screening of any combination of mycotoxins with minimum of three from a list of ten mycotoxins assays available to suit the requirements of testing laboratories. This study presents analytical performance of the technology when animal feed samples were analysed for the most prevalent mycotoxins.Methodology. Simultaneous competitive chemiluminescent immunoassays, defining discrete test sites on the biochip surface and applied to the Evidence Investigator analyser were employed. Assay validation was based on 2002/657/EC. Twenty three animal feed samples from the Food Analysis Performance Assessment Scheme (FAPAS) were analysed. All the mycotoxins were extracted from each feed sample using a single generic liquid/liquid extraction developed in-house. Results. Analysis of ten animal feed samples from FAPAS proficiency testing programme and thirteen FAPAS QC showed values within the range assigned by FAPAS (Z-score ±2) for proficiency testing and recoveries between 79 and 115% for QC samples. Analysed samples presented single or multi-mycotoxin contamination. Conclusions. The results indicate not only optimal performance of BAT for the semi-quantitative multi-mycotoxin screening of animal feed samples based on validation data but also the ability to detect co-occurrence of different mycotoxins by testing FAPAS samples.

P22. Adecisionsupportsystemtocontrolmycotoxincontaminationinmaizesilages

VandickeJ. 1,DebevereS.2,3,FievezV.2,CroubelsS.3,DeSaegerS.4,AudenaertK.1, Haesaert G.. 1

1GhentUniversity,DepartmentofAppliedBiosciences,FacultyofBioscienceEngineering,Ghent,Belgium2GhentUniversity,DepartmentofAnimalProduction,FacultyofBioscienceEngineering,


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Ghent,Belgium3GhentUniversity,DepartmentofPharmacology,ToxicologyandBiochemistry,FacultyofVeterinaryMedicine,Ghent,Belgium4GhentUniversity,DepartmentofBioanalysis,LaboratoryofFoodAnalysis,FacultyofPharmaceuticalSciences,Ghent,Belgium

Mycotoxins are toxic secondary metabolites produced by a variety of fungal species, such as Fusarium, Penicillium or Aspergillus, among others. Contamination of feed with mycotoxins can cause severe health problems in dairy cattle. Especially high yielding dairy cows with a high feed uptake and rapid ruminal flow are susceptible to gastroenteritis, reduced reproduction and reduced milk production, as a result of mycotoxin contamination.Maize silage is one of the main components of dairy feed in the region of Flanders, Belgium, and is therefore one of the main sources for mycotoxin uptake in dairy cows. This research aims towards providing dairy farmers in Flanders with a user-friendly prediction model, able to foresee mycotoxin contamination based on weather, cultivation, harvest and silage conditions. This model will be constructed based on analyses of maize silages across Flanders, and on own research focusing on methods to prevent mycotoxin contamination. 100 maize silages will be selected based upon geographical spread, cultivation technique and silage conditions. These silages will be sampled once during harvest and 2-3 times during feeding every year for four years, and analyzed for mycotoxin and fungal contamination. Own research will be divided into five separate work packages, with the following topics: biofumigation of the soil using green crop manures, treatment of crop residues with antagonistic microbial populations, impact of harvest date and dry matter content on mycotoxin contamination, microbial detoxification in the silage, and toxicity of mycotoxins in dairy cattle. These results will aid towards constructing and validating the prediction model.

P23. Transferofmercuryfromfeedtofish

AmlundH.,LundebyeA.-K.,RasingerJ.D.andBerntssenM.H.G.National Institute of Nutrition and Seafood Research, NIFES, Norway

In Europe, the levels of contaminants in feed and food, including seafood, are controlled through the European feed and food legislation, which set statutory limits for a wide range of contaminants in feed ingredients, feed and food. Maximum levels are established in order to control the introduction of contaminants into the food production chain. However, the maximum limits are often set based on background levels, and do not always take into account the potential transfer of contaminants from feed to food. The European Food Safety Authority (EFSA, 2008) stated that the current maximum level for mercury in fish feed is sufficient to ensure a mercury level in farmed salmon fillet that does not pose a risk to

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consumers. This statement was based on occurrence data and not data on the transfer of mercury from feed to fillet. The EFSA also states that the validity of the maximum level needs to be ascertained for other species of farmed fish.

In a series of trials we have investigated the assimilation and depuration of dietary methylmercury (MeHg) in farmed fish (Atlantic cod and Atlantic salmon), as well as in a model organism (zebrafish). In general, fish readily take up MeHg from feed and MeHg is efficiently accumulated in the muscle, where it is incorporated into larger peptides and/or proteins. A continuous accumulation of mercury in muscle was seen during the exposure periods, and the elimination of mercury from muscle was slow and incomplete within the applied depuration periods. The transfer of MeHg, described by the estimated absorption efficiency, from feed to fish was assessed, and for Atlantic salmon a predictive model for the transfer was developed based on uptake and elimination kinetics. We have also investigated the influence of dietary selenium on MeHg bioaccumulation. Data indicate that selenium reduced accumulation and enhanced elimination of MeHg. An overview of these studies will be given, and the implications for seafood safety will be discussed.

P24. Ethoxyquininfishfeedandfarmedfish

SandenM.,LieØ.&LundebyeA.-K.National Institute of Nutrition and Seafood Research, NIFES, Norway

Storage of animal feed may lead to lipid peroxidation through auto-oxidation. Oxidation of lipids can result in a decrease of nutritional value, an increase in undesired oxidation products and a risk of self-ignition during sea transport of e.g. fish meal over long distances. The antioxidant ethoxyquin (EQ, E324) is one of a number of authorised synthetic antioxidants that is commonly added to fishmeal in order to prevent it from becoming rancid. However, the use of EQ in feed results in residues of this compound and its metabolites in e.g. fish muscle. EQ is currently being assessed for reauthorisation in the European Union. The mean concentrations of EQ in Norwegian fish feeds in the years 2003-2016 varied between 6 and 50 mg EQ/kg but with some feeds exceeding the maximum limit of 150 mg/kg. Most antioxidants act in a self-sacrificial manner leading to the formation of degradation products. The stability of EQ in both fish meal and fish feed was investigated and showed little accumulation of oxidation products such as the ethoxyquin dimer (EQDM) during 250 days of storage. However, the use of EQ in feed leads to residues of both parent compound and oxidation products in foods of animal origin. The residues of EQ and the metabolites EQDM and de-ethylated EQ in species of farmed fish were investigated and showed highest accumulation in Atlantic salmon, followed by rainbow trout, Atlantic halibut and Atlantic cod. The proportion of parent EQ compared to the sum of EQ and EQDM in fillets from commercially farmed fish varied depending on the species. In Atlantic salmon, EQDM is a


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major oxidation product and analysis of farmed salmon in the period 2006-2014 showed that parent EQ was less than 10% of the sum of EQDM and EQ. However, the concentrations of EQ and EQDM in fish have declined in recent years. Analysis of EQ is difficult due to the range of oxidation products that can be formed and some methodological challenges will be presented.

P25. DevelopmentofaCENstandardontheLC-MS/MSanalysisofergotandtropanealkaloidsinanimalfeed.Resultsofacollaborativetrial.

MulderP.,Pereboom-deFauwD.,deJongJ.RIKILT - Wageningen UR, The Netherlands

Ergot alkaloids (EAs) are the toxic principles of rye ergot and tropane alkaloids (TAs) are the toxicants present in Datura seeds. Rye ergot from Claviceps sp. and seeds from Datura sp. are listed as botanical impurities for which maximum limits have been set in legislation (Directive 2002/32/EC). No limits have been established for the alkaloids themselves, but the European Commission recommends that the presence of EAs should be monitored in feed and food (2012/154/EC). Ergocornine, ergocristine, ergocryptine, ergometrine, ergosine and ergotamine and their epimeric –inine forms are considered the major constituents of ergot, while atropine (hyoscyamine) and scopolamine are the predominant TAs in Datura.

In 2013 the EC issued a tender for a new mandate on standardisation of analytical methods for, amongst others, contaminants, mycotoxins, plant toxins and feed additives in animal feed. EAs and TAs are included in this mandate as two separate items (Mandate 522/ items 1 and 2) and with the specific requirements that the analytical method should be LC-MS/MS and cover the alkaloids mentioned above. The LOQ for the individual compounds should be 10 µg/kg. RIKILT has an in-house validated method available that includes the 12 EAs as well as the 2 TAs. It was thus proposed to combine the two items into a single method. This proposal was selected and granted by the EC.

The envisioned standard is being prepared within CEN/TC 327 – Animal feeding stuffs – Methods of sampling and analysis. Work started in May 2014 and the project has a run time of 4 years. The preparation of suitable materials was particularly challenging, but finally homogeneous materials could be obtained. Laboratories were invited to participate early in 2016 and a pre-trial sample has been sent out to familiarise labs with the method. Start of the full trial is scheduled for beginning of June and results are due in August. The results of the collaborative trial will be presented and discussed.

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P26. Newstandardizedmethodsforthedeterminationofnon-dioxin-likePCBs and OCPs in feed

vanLeeuwenS.P.J.,vanderWegG.,ElbersI.,deJongJ.RIKILT - Wageningen UR, The Netherlands

Analytical methods for determination of contaminants such as non-dioxin-like (ndl) polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) are crucial in the control of feed and feed ingredients. Compliance with legislation relies on analytical determination of these contaminants in feed(ingredients). To support laboratories worldwide on this task, CEN/TC 327 published two standards in 2009, being EN 15742:2009 (GC-ECD) and EN-15741:2009 (GC-MS). However, a need for adaptation of these standards rose due to the installation of legislation for ndl-PCBs and the need for inclusion of additional OCPs. More specifically it concerned: (1) extending the scope of the methods for photoheptachlor, cis/trans nonachlor and keto-endrin and (2) reduction of the LOQ for the sum of ndl-PCBs from 5 ng/g per congener to 0.5 ng/gram.The current standards (EN 15742:2009 (GC-ECD) and EN-15741:2009 (GC-MS)) were modified and in-house tests demonstrated that the modified standards were fit for purpose. The modifications of EN 15742:2009 (GC-ECD) concern the inclusion of above mentioned additional pesticides. In addition, the determination of the ndl-PCBs is removed from this standard as GC-ECD is not capable of reaching the lowered LOQs for the ndl-PCBs. Therefore, the modified EN 15742:2009 (GC-ECD) limits to OCPs only. The modifications of EN 15741:2009 (GC-MS) concern the inclusion of above mentioned additional pesticides, as well as the lowered LOQs for the ndl-PCBs. The latter is achieved by either a 10-fold higher sample intake combined with GC-MS or the use of GC - triple quadrupole mass spectrometry (MS/MS). These modifications were reflected upon with a small group of laboratories in order to check of the protocol was clear and experimentally sound. To that end, these labs analysed a small number of samples.Finally, the validation of the modified standards took place in a collaborative trial. Laboratories from around the globe were invited to participate. Eight laboratories subscribed for the collaborative trial on the modified EN 15742:2009 (GC-ECD) standard and 18 laboratories subscribed for the collaborative trial on the modified EN 15741:2009 (GC-MS) standard. The results of both collaborative trials will be presented. These modified standards will, once available, continue to support the production of safe feed in Europe and worldwide.


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P27. DetectionofruminantDNAinfishfeedingstuffscontainingprocessedanimalprotein(PAP).

Desiato R.,GiovanniniT.,BenedettoA.,AmatoG.,CeppaL.,GrifoniF.,RuG.,MarchisD.Istituto Zooprofilattico Sperimentale PLVA, Torino, Italy

Objectives. In the frame of the risk management of transmissible spongiform encephalopathies, recently in the EU legislation processed animal proteins (PAP) from non-ruminants or feedingstuffs which contain them have been reauthorised for feeding of aquaculture animals. Our aim was to describe the pattern of contamination from ruminant DNA in fish feedingstuffs containing PAP collected in Italy over 2015. Eighty-one feed samples intended for fish feeding were collected in the frame of the National monitoring plan of feedingstuffs and submitted to real-time PCR assay to ensure that the PAP originated exclusively from non-ruminants. Part of the samples (74) were from the national random monitoring, part (7) from a risk-based monitoring at the Italian border. The samples were from the following feed categories: complete feed for fish (63 samples; milk was declared in label in 8 of them); raw materials (18 samples: 1 feather meal, 1 blood meal, 3 pork meals, 13 non specified PAP). Results and Conclusion.The seven non specified PAP sampled in the frame of the risk-based sampling were all positive. When considering the random monitoring, that provide a representative snapshot of the Italian situation, the positive tested samples were 22 out of 74 i.e. 29.7%; (95%CI 19.7 – 41.5). In particular ruminant DNA was detected in 17 complete fish feedingtuffs i.e. 27.0%; (95%CI 16.6- 39.7; milk was in label in 1 case), and in 5 out of 11 raw materials i.e. 45.5% (95%CI 16.7 -76.6). The feather meal and the pork meals were positive. The risk-based surveillance was very efficient in detecting the positive raw materials. Moreover the results of the random monitoring raises a concern on feed safety, likely linked to cross-contamination problems. Despite the overlapping of the confidence intervals, a higher proportion raw materials seem to be contaminated when compared to complete feeds containing PAP: raw materials contaminated with ruminant DNA pose a risk also for farmed species other than aquaculture animals.

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P28. Acomprehensiveapproachforthedetectionofregulatedandun-regulateddioxinstoinvestigatetheirphysicochemicalbehaviorintheprocessingofpalmoilandpalmfattyaciddistillate

HoogenboomLAP1,Taverne-VeldhuizenWilma2,3,vanLeeuwenSPJ11 RIKILT Wageningen University and Research, Wageningen, The Netherlands 2 Wageningen UR - Food Quality and Design, Wageningen, the Netherlands3 Cargill Rotterdam, Rotterdam

ObjectiveSeveral non-compliant palm fatty acid products [1] raised the question whether the un-regulated and specifically higher chlorinated dioxins could transform to lower chlorinated dioxins during the processing, among which the more toxic 17 regulated WHO-dioxin congeners [2]. To answer this question an analytical method was developed to determine concentrations of as many dioxin congeners as possible.

Materials and MethodThe standard operating procedure [3] was adapted to include also un-regulated dioxins. The adaptations included, (i) addition of more congeners to the method (Comprehensive Polychlorinated Dioxin and Furan Column Defining Kit, CIL-JR-PCDD/F-KIT and several individual standards of lower chlorinated dioxins) and 13C-labeled internal standards for lower chlorinated dioxins while for unavailable congeners a numerical peak allocation was applied, (ii) addition of ion traces for the lower chlorinated dioxins and acquisition window time adjustments

Results and ConclusionUsing the extended and optimized method we were able to determine all WHO-dioxins and nearly all other PCDD/F congeners. However, some congeners could not be determined or could not uniquely be identified due to interfering or similar retention times. Although not all congeners could be uniquely identified or named, sufficient information was provided to enable the evaluation of the transformation of higher chlorinated dioxins to lower chlorinated dioxins. Using this method the dechlorination of dioxins and the increase in levels of WHO-dioxins during palm oil hydrogenation was demonstrated. A 400% increase of dioxin-TEQ (lb) was found, resulting in non-compliance [4] compared to the original product.

References[1] RASFF, 2014, https://webgate.ec.europa.eu/rasff-window/portal/[2] M. Van den Berg et al., Toxicol Sci 93 (2006) 223[3] R.L.A.P. Hoogenboom et al., Chemosphere 150 (2016) 311[4] Commision regulation (EU) No 744/2012 amending Directive 2002/32/EC


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P29. Developmentofnearinfraredspectroscopymethodsforthedetectionofergotincerealflour

VermeulenP. 1,EbeneM.2,FernándezPiernaJ.A.1, Lecler B. 1,VeysP.1, Baeten V. 1

1 Walloon Agricultural Research Centre (CRA-W), Gembloux, Belgium.2 Université Catholique de Louvain (UCL), Louvain-la-Neuve, Belgium.

Since decades, classical near infrared (NIR) spectroscopy is widely used in the food and feed sectors to implement rapidly, inexpensive and efficient control tools for the quality of products. In the two-thousands, in the framework of the BSE (bovine spongiform encephalopathy) crisis, NIR microscopy (NIRM) and NIR hyperspectral imaging have been widely studied for the authentication of feed ingredients and therefore the detection of animal ingredient particles in feedstuffs [1]. More recently, this technology was investigated to detect undesirable and toxic contaminants in cereals intended for the food and feed sectors. In particular, methods were developed for the on-line detection and quantification of ergot bodies formed by the fungi Claviceps purpurea, which contain toxic alkaloids [2].

The work presented in this poster aims to assess NIR microscopy (in transmission mode) and NIR hyperspectral imaging (in reflection mode) in comparison to classical NIR spectroscopy and classical microscopy to detect particles of ergot bodies in cereal flour. For this study, grounded ergot bodies samples and cereal flour samples from different species (wheat, spelt, rye) and different composition (with or without germ and bran) as well as mixtures of both from 0.01 % to 50 % were analysed. Partial least squares discriminant analysis (PLSDA) models were developed and applied on NIRM mapping or spectral images in order to detect the ergot bodies particles.This research shows the potential of NIRM and NIR hyperspectral imaging combined with chemometrics to propose solutions to discriminate the ergot bodies particles from cereal flour.

AcknowledgementThe authors wish to thank Béatrice Orlando from Arvalis for providing part of the samples analysed in this study.

References [1] Vermeulen, P. , Fernández Pierna, J.A. , Abbas, O. , Dardenne, P. & Baeten, V. (2010). Authentication and traceability of agricultural and food products using vibrational spectroscopy In: Applications of Vibrational Spectroscopy in Food Science, Eunice C.Y. Li-Chan, Peter R. Griffiths, John M. Chalmers. John Wiley & Sons, Ltd, 2, 609-630. [2] Vermeulen P., Fernández Pierna J.A., van Egmond H.P., Zegers J., Dardenne P. & Baeten V. (2013). Validation and transferability study of a method based on near-infrared hyperspectral imaging for the detection and quantification of ergot bodies in cereals. Analytical and Bioanalytical Chemistry, 405: (24), 7765-7772

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P30. EarlydetectionofcontaminantsinfeedmillsbyNearInfraredSpectroscopy and Chemometrics

FernándezPiernaJ.A. 1,AbbasO.1, Lecler B. 1,HogrelP.2,VermeulenP. 1,DardenneP. 1, Baeten V. 1

1 Food and Feed Quality Unit, Valorisation of Agricultural Products Department, Walloon Agricultural Research Center, Gembloux, Belgium.2 Provimi SA, Crevin, France

Objective-The goal of this study is to develop a complete procedure based on Near Infrared (NIR) spectroscopy and chemometrics for characterizing typical feed ingredients and detecting the possible presence of contaminants from various origins directly at the entrance of the feed mill, preventing the contamination of the whole production chain. This study aims avoiding crises such as the recent melamine scandals in pet and human food commodities.Material and Mathods-The procedure proposed includes the use of vibrational spectroscopy and Chemometrics. Vibrational spectroscopy is used as accurate, rapid and sensitive analytical method needed for the assessment of the quality and safety of food and feed products. Chemometrics is needed for the creation of decision rules based on specifications. In this study, a chemometric approach based on a moving window along the wavelength range is proposed for the data-driven discovery or untargeted analysis. For each selected window in a calibration set, a PCA analysis is performed by fixing the number of principal components. Spectral score residuals in the calibration set are extracted and used to build thresholds. When a residual, at a certain wavelength, falls out the defined thresholds, the sample could be suspected of being abnormal and then indicating the possible presence of contaminants or mislabeling. Results and Conclusions-The complete procedure has been implemented in a large feed mill (Cargill Animal Nutrition) in order to detect anomalies due to an accidental or fraudulent addition of contaminant. All the efforts have been put on the setting of multivariate specifications for the NIR spectral characterization of pure soybean meal. The methodology has been tested with different trucks containing the soybean meal that were deliberated contaminated during the loading at the entrance of the production chain. The results demonstrated the possibility to implement fast and efficient control strategy to detect on-line contamination (whatever the origin) at the discarding place. This work has been funded by the European Commission’s Quality and Safety of Feeds and Food for Europe (QSAFFE) – 265702: Collaborative Project under the Seventh Framework Programme, KBBE-2010-2-4-03: Quality and Safety Aspects of Feed.


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P31. Studyonhighmarkedspectralpropertiesforspeciesidentificationofanimaloriginatedfeedbyft-irbasedonlipidcharacteristics

LiuX.,GaoF.,Han L.College of Engineering, China Agricultural University, Beijing, China

There has been increased attention towards the fraud of animal originated feed which play an important role in the animal husbandry industry. Considering the high security risk of animal originated feed, effective quality control and reliable discriminant method is becoming more and more important. The objective of this study was to explore the capability of FT-IR for discriminating different species of animal-originated feed based on the lipid characteristics, and figure out high marked spectral regions (single or combined) which have main contribution to the species discrimination. A total of 85 reliable MBM (including porcine, poultry, bovine, ovine) and FM samples were involved in this study and their fatty acid profile was analyzed by GC. The raw materials, extracted lipid and defatted samples were simultaneously analyzed by FT-IR in combination with chemometric methods. Results showed that successful discriminations by FT-IR were achieved among the tested five species of animal-originated feed based on lipid characteristics, whose values of sensitivity and specificity were very closed to 1. The correlation between lipid properties and infrared spectral characteristics were further analyzed, and five spectral regions were considered as the high marked spectral properties for species identification. Discriminant analysis models of marked spectral bands were investigated and proved to have better performance than the whole spectra. For five species discrimination, the combined regions of 1800-1650 cm-1+1500-1330 cm-1 + 1260-1060 cm-1 + 790-640 cm-1 presented better result; while for three category classification (non-ruminant MBM, ruminant MBM and FM), combined regions of 3100-2800 cm-1 + 1800-1650 cm-1 + 1500-1330 cm-1 showed the best result. It is possible to consider FT-IR spectroscopy as the valuable and powerful analytical methodology for the rapid discrimination of animal-originated feed, especially based on the lipid characteristics.

P32. RapidMultiresidueandMulti-classanalysisforAntibioticsandAnthelminticsinFeedbyUltrahighperformanceliquidchromatographycoupled to tandem mass spectrometry

RobertC.,BrasseurP.Y.,DuboisM.,GillardN.CER Groupe, Health Department, Marloie, Belgium

Conventional livestock production systems use antibiotics therapeutically, prophylactically, and as growth enhancers. The presence of antibiotics in feed is either authorized (for

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therapeutic and prophylactic purposes), unauthorized (antibiotics as growth promoters), or unintentional (due to cross-contamination).The authorized antimicrobials most broadly used in medicated feed are tetracyclines, sulfonamides, trimethoprim, macrolides, blactams, aminoglycosides, pleuromutilins, and lincosamide. The use of medicated feeds is most common in intensive production, especially of pig and chicken (European Commission, 2010).Although these antimicrobials are authorized, traces are undesirable in non-medicated feed. As medicated and non-medicated feeds are often manufactured in the same production line, carryover of antimicrobials can occur when a feed miller switches from producing one feed to the next (Stolker et al., 2013) or later in the production line.An analytical strategy was developed for high-throughput screening of multiple antibiotics and two benzimidazoles in feed. Generic sample processing was applied without any purification step. After methanol extraction, the samples were centrifuged, concentrated, and analysed by ultra-highperformance liquid chromatography hyphenated to tandem mass spectrometry in the multiple reaction monitoring mode. Qualitative validation was carried out for more than 54 antibacterials of various classes, including aminocoumarin, amphenicols, beta-lactams, lincosamide, macrolides, diaminopyrimidine, nitrofurans, quinolones, sulfonamides, streptogramin, pleuromutilin, polypeptide, quinoxaline, and tetracyclines, and also some benzimidazoles in feed at µg/kg level. Validation was done in accordance with the guidelines laid down in European Commission Decision 2002/657/CE for qualitative screening methods.This convenient, reliable, and sensitive method has been used successfully to monitor antibiotic residues in feeds.

P33. Comparisonofvariouschromatographicconditionsandmassspectrometers for the analysis of coccidiostats in feed

Gerritsen H. 1,ZuidemaT.1, Vincent U2

1 RIKILT - Wageningen UR, The Netherlands, 2 European Commission, Directorate General Joint Research Centre, Geel, Belgium

From the late 1960s coccidiostats are in use to combat the spread of coccidiosis, particularly in the poultry sector. The first used coccidiostats were the naturally produced by bacteria Streptomyces spp. and Actinomadura spp [1], namely lasalocid, monensin, salinomycin, maduramicin, narasin and semduramicin, also called the ionophoric coccidiostats. Later also chemical coccidiostats (diclazuril, halofuginone, decoquinate, nicarbazin and robenidine) were developed for treatment against coccidiosis [2]. Coccidiostats are the only anti-bacterial substances still authorised and registered as feed additives for chickens, turkeys, rearing laying hens and rabbits. In the production of compound feeding stuffs using these additives, carry-over may occur to the next batch compound feeding stuffs. In directive 2009/8/EC [3],


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maximum levels are stated for unavoidable carry-over of coccidiostats and histomonostats to non-target feed. The LC-MS/MS analysis of coccidiostats, in particular the ionophoric ones, is often hindered during LC-MS/MS analysis. Ion suppression seems to be one of the main causes. In this study and with the aim of assessing the transferability of a candidate standard method for the determination of coccidiostats, different types of mass spectrometers were used to examine differences between instruments and the instrumental impact on the analysis of these analytes. Furthermore, the use of HPLC versus UPLC was also examined. Measurements were executed on two different triple quadrupole mass spectrometers: QTRAP5500 (Sciex), and XEVO-TQS (Waters) and on a high resolution mass spectrometer: Q-Exactive (Thermo Scientific). Results of the comparison will be presented on a poster.

P34. Dumpfromillegaldruglaboratories–anewriskforanimalfeed?

ZuidemaT.,Lasaroms,J.J.P.,deJong,J.RIKILT - Wageningen UR, The Netherlands

Illegal production of XTC and synthetic drugs is increasing rapidly. Laboratories expand and professionalize. In the Netherlands not only 20 to 30 laboratories are tracked down yearly, but also the same amount of illegal dump spots are uncovered. Until now most of these dump spots are located in nature reserves and agricultural areas. Recently however, waste from XTC-production was found in a manure pit. Whereas the risk of dumping waste from illegal drug production in nature reserves and agricultural areas “only” concerns the possibility of contaminating the soil or groundwater, the spreading of contaminated manure on agricultural grounds may lead to drug (or metabolites of drugs) residues in crops and possibly in edible tissues of the animals. These new risks will be presented and discussed.

P35. Adatabasefordioxincongenerprofilesforidentificationofsources

HoogenboomR.L.A.P.1, Malisch R. 2,VanLeeuwenS.P.J.1, Ten Dam G. 1,vanRaamsdonkL.W.D.1, VanderperrenH.3,HoveH.4,FernandesA.5, Schächtele A. 2, Rose M. 5

1 RIKILT Wageningen UR, Wageningen, The Netherlands, 2 CVUA, Freiburg, Germany, 3 FLVV, Tervuren, Belgium4 NIFES, Bergen, Norway, 5 FERA, York, United Kingdom

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ObjectivesPolychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), and dioxin-like PCBs remain a major threat in the food chain, since exposure of consumers is still around the existing health based guidance values. Both self-control by companies and official control by authorities are important to further reduce and monitor existing levels of these contaminants. In the case of samples exceeding action or maximum levels, follow-up studies are required to identify and eliminate the source of the contamination. Congener profiles of PCDD/Fs are important tools in this process. A working group within the EURL/NRL network dioxins was established to collect the various patterns and make them publically available.

Materials and methodsWithin the network profiles of incidents were collected and checked for completeness. This included a number of incidents reported in literature. The initial focus was on primary patterns of PCDD/Fs, i.e. patterns not modified by kinetics and metabolism in food producing animals. An Excel-file was created that allows quick comparison of a new analytical finding with patterns observed in previous incidents.

Results and ConclusionsIn addition to the Excel-file, a decision tree was developed as a first guidance to the source. Major source categories are chlorinated chemicals like PCBs and chlorophenols, certain clay materials and minerals and burning of chlorine containing materials. Patterns containing only PCDFs point to PCBs, patterns with only PCDDs to either clay materials of chlorophenols. Higher chlorinated PCDDs point to pentachlorophenol. Mixed patterns are observed in case of minerals and burning of waste. PCB-patterns are useful to confirm PCBs as a potential source and for determining the type of PCB-mixture causing the incident. Future activities focus on the changes in patterns in farm animals, as well as the possibilities to develop a decision support system.

P36. Candidatestandardmethodforthedeterminationofcarotenoidsinanimalfeedingstuffs

SeranoF.,vonHolstC., Vincent UEuropean Commission, Directorate General Joint Research Centre, Geel, Belgium

Carotenoids are feed additives classified in the category “sensory additives” and functional group “colorants” i.e. substances which, when fed to animals, add colour to food of animal origin (Annex I of Regulation (EC) No1831/2003). Legal limits of these compounds have been established in feed. Moreover, the authorisation of these compounds also includes corresponding Maximum Residue Limits (MRLs) in food matrices. Improved methods of analysis of carotenoids are therefore required in order to carry out official controls to


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ensure the verification of compliance with feed and food law, labelling claims, animal health and animal welfare rules (Regulation (EC) No 882/2004). Carotenoids present a variety of stereoisomers, due to the numerous conjugated double bonds and the cyclic end groups. The most important forms commonly found are geometrical isomers (E/Z); these are inter-convertible in solution and present different physical properties and in particular their absorption coefficient and the wavelength of maximum light absorbance (λmax) are different.A multi-analyte HPLC method which enables the reliable quantification of authorised carotenoids in fish and poultry feeds and pre-mixtures has been successfully validated in-house. This new method has the important advantage that it is applicable to all the different sources of carotenoids (chemical and natural additives), which until now required the use of product specific methods. The new method involves a simple sample preparation, avoids the use of chlorinated solvents, is based on reverse phase HPLC and provides reliable quantification thanks to the isosbestic concept which makes it fit for the purpose of official control of carotenoids in fish feed. The method has been selected as candidate method for standardization under the third mandate, part III (M/523) of the European Commission to the European standardization body CEN.

References[1] Mitrowska K., Vincent U., von Holst C., J Chromatogr A. 2012 Apr 13; 1233:44-53.[2] Serano F., von Holst C., Vincent U., private communication, EURL-FA annual workshop 2015

P37. Determinationofthe11authorisedcoccidiostatsorhistomonostatsinfeedingstuffsbyliquidchromatography-tandemmassspectrometryatcross-contaminationlevels:ananalyticalchallenge

ChedinM.,EzerskisZ.,vonHolstC.,VincentU European Commission, Directorate General Joint Research Centre, Geel, Belgium

A multi-analyte method for the determination of the 11 authorised coccidiostats or histomonostats (halofuginone hydrobromide, robenidine hydrochloride, nicarbazin, diclazuril, decoquinate, monensin sodium, salinomycin sodium, narasin, lasalocid sodium, semduramicin sodium and maduramicin ammonium alpha) has been developed and single-laboratory validated at cross-contamination concentration levels in feed by liquid chromatography tandem mass spectrometry (LC-MS/MS). The corresponding maximum cross-contamination levels have been recently introduced by European legislation. The method developed involved a simple liquid-solid extraction of the coccidiostats or histomonostats from the feed samples followed by centrifugation and filtration of the supernatants before determination by LC-MS/MS. Quantification was performed using a modified standard additions-based approach, hence allowing a workload comparable to a matrix-matched standard calibration curves approach or even less.

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The method was validated at three concentration levels, namely at half the cross-contamination level, the cross-contamination level and twice the cross-contamination level. The intermediate precision expressed in terms of relative standard deviation varied between 3 and 14 % and the recovery rates ranged from 85 to 118%, depending on the target analyte and matrix. The limits of detection (LOD) and limits of quantification (LOQ) were different for the various analyte/matrix combinations but all LOQs were in the 0.005 mg kg-1 to 0.18 mg kg-1 range and in the 0.02 mg kg-1 to 0.34 mg kg-1 range for poultry and pig compound feed respectively, hence well below the target concentrations of each analyte. Based on the obtained performance characteristics during the single-laboratory validation, the method is considered fit for the purpose of the official control of the 11 authorised coccidiostats in feed at cross-contamination level.

[1] U. Vincent, M. Chedin, S. Yasar, C. von Holst, J. Pharm. Biomed. Anal., 47 (2008) 750 – 757.[2] U. Vincent, Z Ezerskis, M. Chedin, C. von Holst, J. Pharm. Biomed. Anal., 54 (2011) 526 – 534.

P38. Consequences of using feather and/or bone meal as feed ingredient

ZuidemaT.1,vanEgmondH.J.1,vanBrakelM.W.2,PelkM.L.H.2,deJongJ. 1 1 RIKILT Wageningen UR, Wageningen, Netherlands2 NVWA, Utrecht, Netherlands

Veterinary drug residues in animal feed are an issue to consumer, animal and environmental health. Not only for toxicity reasons but also for other impacts including the induction of resistance in target organisms. Besides medicated feeds and deliberate addition of antibiotics for growth-promoting effects, other causes may motivate the presence of antimicrobial residues in feed. These residues may be caused accidentally, such as through cross-contamination of non-medicated feed in facilities producing medicated feeds, or through the inclusion of contaminated feed ingredients, for example from animals or plants treated with antimicrobials. As feathers and bones tend to store antibiotics (and possibly other contaminants) feather meal and bone meal should be considered high risk feed ingredients. Even hydrolysed materials may contain contaminants such as antibiotics. The risk of using feather meal and bone meal as feed materials will be discussed.


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P39. Occurrence of ergot alkaloids in cereals from Albania

Tavčar-KalcherG.,Jakovac-StrajnB.,TopiD.University of Ljubljana, Veterinary Faculty, Ljubljana, Slovenia

Ergot alkaloids (EA) are secondary metabolites produced mainly by fungi of Claviceps genus. They are mostly present in cereals and wild grasses. A maximum level of 1000 mg/kg of rye ergot (Claviceps purpurea) sclerotia has been established for feed containing unground cereals. However, European Commission recommended Member States to perform monitoring on the presence of ergot alkaloids in cereals intended for human consumption or animal feeding (Commission Recommendation 2012/154/EU). This study is the first reporting the ergot alkaloid presence in cereals harvested in Albania in two consecutive years.

Wheat, barley and rye were collected in summer 2014 and 2015 in four main producing regions of the country. A total of 42 and 41 cereal samples were analysed in 2014 and 2015, respectively, using liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of most important EA: ergometrine, ergosine, ergocornine, ergocryptine, ergotamine and ergocristine and corresponding -inine epimers.

In 2014, EA were present in 17 out of 35 wheat samples (49 %), in three out of five barley samples (60 %), and were not detected in rye samples. The total alkaloid concentrations were in the range 17–975 µg/kg in wheat and 89–492 µg/kg in barley. In the year 2015, six out of 35 wheat samples (17 %) and one out of three rye samples (33 %) contained EA. None of two barley samples was contaminated. In wheat, the total alkaloid concentrations were in the range 10–390 µg/kg and the concentration in rye was 995 µg/kg.

In 2014, each contaminated wheat sample contained one to nine EA and each contaminated barley sample four to five EA. In 2015, the contaminated wheat samples contained one to six EA, but the contaminated rye contained eleven EA. In 2014, the most frequently present EA were ergometrine and ergosine, while ergocorninine and ergocryptinine occurred least frequently. In 2015, most frequent were ergocristine and ergicristinine.

P40. Cadmium,lead,mercuryandarsenicinanimalfeedandfeedmaterials-trendanalysisofmonitoringresults.

AdamseP.,vanderFels-Klerx,H.J.,deJong,J.RIKILT Wageningen UR, Wageningen, the Netherlands

Data from the National Feed monitoring program of the Netherlands Food and Consumer Product Safety Authority (NVWA), the Product Board Animal Feed and representatives from

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the feed industry have been used to study trends in maximum limit (ML) exceedances and average, median and 90th percentile concentrations of cadmium, lead, mercury and arsenic in feed and feed materials in the Netherlands between 2007 and 2013. The data relate to monitoring research and do not contain results from targeted samples taken as a follow-up on incidents. In most samples all heavy metals were analysed at the same time. Between 300 and 500 samples were analysed each year. The results of these analyses enable the NVWA to improve their risk-directed sampling strategy.

Based on the results, monitoring for cadmium and lead should focus on complementary feed, feed material of mineral origin and feed material of animal origin, especially fish meal. No forage samples have been reported with lead concentrations exceeding the ML. For complementary feed it is important to make a proper distinction between mineral and non-mineral feed because the ML for lead in the latter group is lower. For feed material of mineral origin and feed material of animal origin (especially fish meal), an increase was seen in both the percentage of samples exceeding the ML for cadmium and the average cadmium concentration . For mercury, an increasing presence was seen in fish meal and premixtures . The concentration of arsenic increased significantly in feed materials of mineral origin and in complementary mineral feed. In seaweed/algae products a relative large number of samples contained arsenic concentrations that exceeded the ML. However, most samples were from 2009, which made it impossible to determine trends.

Recommendations: For many feed categories a low sampling frequency is sufficient, but from complementary mineral feed, binders and anti-caking agents and several feed materials of mineral origin more samples should be collected. Both fish meal (cadmium, mercury) and algae (arsenic) should be monitored as well.

P41. Standardisationofamethodforthe“DeterminationofmelamineandcyanuricacidinanimalfeedbyLC-MS/MS”

FryH.,MietleK.,MähnertE.,ZinkeS.,SchwietersM.,PyddeE.,Preiß-WeigertA.Bundesinstitut für Risikobewertung (BfR), department contaminants, Berlin, Germany

To pretend high protein contents melamine and cyanuric acid were illegally mixed into food and feed. As a consequence for melamine a maximum level of 2.5 mg kg-1 feed was established under directive 2002/32/EC. For the control of the maximum level a method to quantify melamine and cyanuric acid is required. Therefore the Technical Committee (TC) 327 “Animal feeding stuffs – Methods of sampling and analysis” of the European Committee for Standardisation (CEN) commissioned within the framework of the third mandate the validation of a method for the analysis of melamine and cyanuric acid in animal feed.The method to be validated consists of a screening and a confirmatory part. In the screening


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method the analytes have to be extracted once, diluted and analysed by LC-MS/MS. For samples tested positive a confirmatory method has to be carried out. Therefore analytes have to be extracted three times, after addition of internal standards, two dilution steps and a standard addition for quantification purposes has to be performed. The LC-MS/MS conditions included in the method are recommended but not mandatory. The method is in-house validated with a limit of detection and a limit of quantification below 1 mg kg-1.For further method validation a collaborative trial is necessary. In a pre-trial four training samples were sent to 12 participants to get familiar with the method. Only 7 laboratories reported results, which demanded a further call for participation, leading finally to 13 laboratories in the main study.Eight feed samples with different concentrations of melamine and/or cyanuric acid were prepared. After homogeneity testing they were distributed as double blind samples subsequently to the participants. Additionally standards, internal standards and a declared blank material were sent. For performance control the newly participating laboratories received also samples from the training period with specified concentrations.

P42. Determinatorastoolforidentifyingdioxinprofiles

RaamsdonkL.W.D.van,HoogenboomL.A.P.Wageningen UR, The Netherlands

Identification is an implicit part of all analytical techniques. In a range of cases libraries with reference objects are in use. Examples of libraries are the GC/MS library NIST14 and Genbank with DNA sequence information.In the discipline of biology, as in many other areas, expert systems are being used for the identification of a range of different entities. Determinator is a platform for application of expert systems, developed by Wageningen UR, and applied in the field of feed and food safety. The core of the system is a matrix with decision rules, each specifying the relationship between a characteristic feature of a subject and the state of that feature in a reference object from a library. This principle can be applied to identification of chemical profiles as well.

A basic profile resulting from a dioxin analysis usually consists of contamination values for 17 congeners of dibenzo-p-dioxins and dibenzofurans, and 12 priority congeners of biphenyls. These 29 values can be normalised in order to get mutual comparability with other profiles. The value of each congener is considered a single characteristic feature, making a total of 29 features describing the subject profile originating e.g. from an incidence.In the current case a library of normalised dioxin reference profiles was developed in Determinator. Three subject profiles were used for identification in order to present the proof of concept. The procedure for identification an incidence profile by means of Determinator

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includes two steps: (a) the input of the normalised shares of all the congeners of the profile and (b) calculating the matches between this incidence profile and all reference profiles. In all cases a relevant match was found. Several modifications of the strategy have been tested.The dioxin module in Determinator provides further documentation for pinpointing the incidence profile. In the view of a frequent occurrence of dioxin incidences, it is highly desired to support the authorities by finding the possible cause and origin of the contamination. The advantage of a formalised system for identification of dioxin profiles is the reproducibility of the conclusion, easy distribution of approved expertise and a levelled playing field for all users.

P43. Howdonationalreferencelaboratoriesperformwhenanalysingheavymetalsinfeed,since2006?

FiamegkosI.,DelaCalleB.,DehouckP.,CordeiroF.,RobouchP.European Commission, Directorate General Joint Research Centre, European Union Reference Laboratory for Heavy Metals (EURL-HM), Geel, Belgium Regulation (EC) No 882/2004 attributed to the Joint Research Centre (JRC) Geel of the European Commission (EC) the management of the European Union Reference Laboratory for Heavy Metals in Food and Feed (EURL-HM).

One of the main mandates of the EURL-HM is to organise proficiency tests (PTs) to assess the performance of National Reference Laboratories (NRLs) in the determination of heavy metals in food or feed commodities. A total of twenty three PTs were organised since 2006.

This poster will review the performance of NRLs for the determination of arsenic, cadmium, mercury and lead in various feed samples. An improvement in performance was observed throughout the years, mainly due to the exchange of good practices among the members of the network. However, several matrices - such as mineral feeds or clays - were found to be “challenging” to analyse, and arsenic is the analyte with “room for improvement”.

P44. Combinedscreeningofcontaminants,undesirablesubstances,andresiduesinfeedbyliquidchromatographywithhighresolutionmassspectrometry

ZomerP.,BorG.,Wegh,R.S.,ZuidemaT.,Mol J.G.J.RIKILT Wageningen UR, v, The Netherlands


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Feed and feed ingredients comprise an extremely wide variety of matrices ranging from wheat, by-products from food and biofuel industry, to complete feeds. During production, storage and processing, a wide variety of substances may enter the feed chain which might affect animal health/production, or safety of animal products for human consumption. Examples are environmental contaminants, mycotoxins, plant toxins, residues of pesticides and residues of veterinary drugs. Dedicated methods exist for prioritized and/or regulated compound/matrix combinations but these are far from comprehensive and changes are high that undesirable substances may go unnoticed.

The aim of this work was to develop a chemical screening method capable of detection of a very wide variety of substances in representative feed materials. For this, existing methods and approaches were (re)considered to select the optimum method (best compromise) to achieve a comprehensive screening method. Extraction was based on a mixture of acetonitril, methanol and formic acid. Analysis of the extracts was performed using liquid chromatography with high resolution mass spectrometry (LC-Q-Orbitrap). Full scan acquisition was done using multiple scan events (full scan / vDIA) to have a truly non-targeted measurement while still obtained data on precursor ions and fragment ions which facilitates selective detection and (tentative) identification. In addition, this allows retrospective data analysis for analytes that were unknown or not of interest at the time of analysis.

The applicability and reliability of the method at the relevant limits of detection were evaluated and will be presented. The method is fairly straightforward but does require analysis in a more advanced laboratory setting. On the other hand, the method is second to none with respect to coverage of numbers of potentially hazardous compounds and very suited to detected substances beyond the known usual substance/matrix combinations.

P45. Applicationofgaschromatography-electronionization-fullscanhighresolutionOrbitrapmassspectrometryfordeterminationofundesirablesubstances in feed

TienstraM.,ZomerP.,Mol H.RIKILT Wageningen UR, v, The Netherlands

High resolution mass spectrometry (HRMS) is an emerging technology in the field of residue and contaminant analysis. In full scan mode, a non-targeted measurement is performed which is more straightforward and better suited for simultaneous detection of very high numbers of analytes than triple quadrupole MS. While in liquid chromatography (LC) the application of HRMS has matured over the past decade and the technique can now be considered suited for routine application, developments in gas chromatography (GC) have been much slower. Recently, a GC-EI-Q-Orbitrap MS instrument has become available offering mass

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resolving powers up to 120,000, which is much higher than other GC-HRMS instruments and considered of high potential for feed analysis. Applicability of the instrument has been reported for determination of pesticides in fruit and vegetables [1].

The aim of this work was to evaluate the applicability of this new technology for feed and feed materials, which are more complex matrices than fruit and vegetables. Target analytes were undesirable substances from CR 2002/32 (section IV: organochlorine compounds and section V: non dioxin-like PCBs) plus a wide range of pesticides regulated under CR 396/2005. Based on previous work [1], measurements were performed at a resolving power of 60,000 (m/z 200 FWHM). Performance parameters studied included selectivity, limits of detection, quantification and limits of identification. Besides acquisition in full scan mode, the option and potential advantages of combined full scan and selected ion monitoring (SIM) was also studied. The final method was validated and a comparison with an existing method based on GC-EI-MS/MS (triple quad) was made. [1] Hans G.J. Mol, Marc Tienstra, Paul Zomer, Evaluation of gas chromatography - electron ionization - full scan high resolution Orbitrap mass spectrometry for pesticide residue analysis, Analytica Chimica Acta (2016),

P46. Proficiency-testingschemeforfeed

MESSINEO E. 1,DIORÉE.1,LEMONNIERL.1, BOUBETRA A.1

1 BIPEA, PARIS - FRANCE.

BIPEA (http://www.bipea.org) organizes regular proficiency-testing schemes (PTS) in many analytical domains, including physico-chemical analyses of feed. As an example, in September 2015, one test was conducted using feed for piglet matrix, with about 30 to 117 participating laboratories according to the parameters. This test was intended for physico-chemical determinations, using standardized methods. Participating laboratories were required to return their results on a dedicated website after a period of one month, and a statistical treatment of the data was as usual performed by BIPEA according to ISO 13528 [1]. Assigned (consensus) values were calculated from the participants’ results and the performances of the laboratories could then be evaluated individually and collectively according to ISO 17043 [2]. These tests allow participating laboratories to draw up a general inventory of their analytical skills, and are a very useful tool to detect bias or non-compliant results; they act as a warning signal for the implementation of corrective and/or curative actions in the laboratory.


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References

[1] ISO 13528:2015,Statisticalmethodsforuseinproficiencytestingbyinterlaboratorycomparisons, 2015.[2] ISO 17043:2010,Conformityassessment--Generalrequirementsforproficiencytesting,2010.

P47. TheefficiencyofcommercialpreparationsoforganicacidsforeliminationofnaturallyoccurringSalmonellainfeedmaterials

AdlerA.,KolarV.,StrnadI.Austrian Agency for Health and Food Safety, Institute for Animal Nutrition and Feed, Linz, Austria

IntroductionSalmonella contamination in feed is still a serious issue in livestock (esp. poultry). This is caused chiefly by contaminated protein feed materials such as soybean meal and expeller from soya, sunflower and rape seed, or corn gluten. A decontamination of the affected feed with organic acids is already being applied in practice, but often the desired results were not achieved. Against this background, the efficiency of the decontamination of heavily Salmonella contaminated feed materials was tested with commercial organic acid preparations. All feed materials used in the experiments were already on the feed market and had a natural, high-grade contamination with Salmonella in tenfold analysis (10/10).

Material and MethodThe experiments included 3 feed materials (soybean meal, coarsely ground Marian thistle seeds, corn gluten), 5 commercial acid preparations (selected and provided by the manufacturers), 7 concentration rates (1%, 2%, .. up to 7% inclusion of the acid mixture) and 3 exposure times (1, 2 or 7 days storage at room temperature). This resulted in an experimental design with a total of 315 test samples (= 3 x 5 x 7 x 3), each of which tested by 10-fold analysis. Additionally control samples without adding acid were analyzed. Salmonella analyses were carried out according to ISO 6579.

ResultsA successful elimination of Salmonella-positive feed under the given test conditions was not possible with all acid preparations and if so, only could be reached with relatively high acid concentrations: A complete decontamination (0/10) of all three feed materials tested was achieved only with 6% of the most effective preparation after 7 days exposure or with 7% of the same product already after 1 day respectively.

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ConclusionsDue to the high acid concentrations necessary for feed decontamination the use of organic acids for sanitization is only recommended for the treatment of Salmonella-containing feed materials, but not for compound feed and particularly not for finished/complete feed (e.g. due to acid-induced feed refusal and/or interactions with other feed ingredients). Future recommendations for the application of acid mixtures in feed should better distinguish between the decontamination effect in the feed itself and a physiological effect in the digestive tract of the animals.


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POSTERS SESSION 4

P48. effectoforganictracemineralongrowthresponseofbroiler

JunlaphoW. 1,2,PuangmaleeT.2,SongsermO. 1,AttamangkuneS.1,RuangpanitY.. 1

1 Kasetsart University, Kamphaeng Saen Campus, Department of Animal Sciences, Faculty of Agriculture at Kamphaeng Saen, Nakhon pathom, Thailand, 2 Kasetsart University, Kamphaeng Saen Campus, Center for Agricultural Biotechnology, Nakhon Pathom, Thailand

Trace mineral proteinates (peptides) for rapid absorption and assimilation can replace inorganic supplements in broiler diets, possibly lowering inclusion rates and mineral excretions. The objective of this study is to compare the effect of a standard inclusion level of inorganic trace minerals to organic trace minerals at different inclusion levels on broiler performance and meat quality. Organic minerals, with di- and tripeptides, were compared at 3 levels to inorganic minerals 100% (CON). The CON treatment had 40.00 ppm Fe, 8.0 ppm Cu, 60 ppm Zn, 80 ppm Mn, 0.5 ppm I and 0.100 ppm Se added, and organic minerals were added 500g per ton in CON treatment diet (ORG500g+CON), and organic minerals were added 750g per ton in corn soy based diets without inorganic minerals (ORG750g), and other proportions of organic minerals were added 750g per ton and plus organic selenium in corn soy based diets without inorganic minerals (ORG750g+Se). Ross 308 broiler chicks (800 totals) were divided into 4 dietary treatments. Each treatment consisted of 10 replications with 20 boilers per replication (20 males). All birds were raised in environmental controlled house for 49 days. Compared with CON, ORG500g+CON improved average daily gain and average body weight gain from 0 to 17 d (P<0.05) and ORG750g treatment improved average daily gain and average body weight gain from 18 to 49 d (P>0.05). The ORG750g increased 49-dBW compared with CON. Feed intake was higher for ORG500g+CON compared with CON from 0 to 17 d and higher for ORG750g compared with CON from 18 to 49 d. From 0 to 49 d, FCR and mortality percentage did not differ significantly among any of the treatments. The use of organic trace mineral proteinates at moderate higher levels (500g and 750g per ton based diet) during the 0-17d and 18-49d of broiler production led to better performance during this period.

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P49. Effectsofdietarygrapeseedextractongrowthperformanceandfilletbiochemicalcompositionofrainbow trout (Oncorhyncus mykiss)

KesbiçO.S. 1,2,YiğitM.31 Kestamonu University Inebolu Vocational School, Kastamonu, Turkey;2 Çanakkale 18 Mart University, Graduate School of Natural andApplied Sciences, Çanakkele, Turkey; 3 Çanakkale 18 Mart University, Aquatic Sciences and Technology Faculty, Çanakkel, Turkey.

IntroductionThe use of antibiotics as growth promoters in feed for fish is banned in many countries because of potential effects on humans and fish. Grape seed extract (GSE) may serve as an alternative feed additive for growth promotion in fish. Therefore the present study was aimed to examine the effects of different levels of grape seed extract as a feed additive on growth performance and fillet biochemical composition of rainbow trout.

Material and MethodsThe rainbow trout (7.35±1.88 g) were placed in 12 aquariums and the aquariums volumes are 100L. The mean initial stocking density was 20 fish per aquarium. GSE was added dosage of 0.5, 1 and 2 g kg -1 in commercial trout feed. Fish were fed two times a day ad libitum for 60 days.

Results and ConclusionAll experimental diets were accepted by the fish. The feed supplemented with GSE have impact on fish growth performance and fillet crude protein level. The experimental groups which include 0.5 and 1 g kg-1 GSE showed lower feed conversion ratio value compared with control and 2g kg-1 GSE group. Protein utilization rate is positively effected by 0.5 and 1 g kg-1 groups compared with control and 2 g kg-1 GSE groups.As a result grape seed extract supplementation especially 0.5 and 1 g kg-1 improve growth performance and fillet crude protein level in rainbow trout.

P50. Mixingtimesandwagonloadaffectstotalmixedrationhomogeneityindairy cows

AgazziA.1,TangorraF.1,PilottoA. 1,TosoniP.C.M.2, Tretola M. 1, Costa A. 1

1 Dipartimento di Scienze veterinarie per la salute, la produzione animale e la sicurezza alimentare – VESPA, Università degli studi di Milano, Milan, Italy.2 Animal nutritionist at DY-PRO, Castel Goffredo, Mantova, Italy

The present study was intended to determine the influence of different cutting and mixing


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times on total mixed ration (TMR) homogeneity and particle size distribution, considering different loading levels of the mixing wagon. The TMR particle size distribution was investigated according to three mixing wagon loads (40, 70 and 100%), b) three cutting times (4, 5 and 6 min), and c) three mixing times (4, 5 and 6 min). The supplemented diet (70:30 forage/concentrate ratio) was supplied by a two-screws vertical mixing wagon (maximum nominal load, 21m3). Data were analysed by a Principal Component Analysis (PCA) accounting for TMR chemical composition of collected samples along the alley and the chemical composition of the designed diet. Obtained results showed that the content in the bottom pan (diam.<1.78 mm), gradually increased (P<0.001) along the TMR distribution in the alley. These preliminary results show that smaller particles of concentrates are submitted to demixing during feed release, with potential unbalances in feed administration. Moreover, 100g of the same samples previously dried to 12% humidity, were sieved for 20 min through an electromagnetic sieve shaker (Retsch VS 1000), with sieves arranged from the largest to the smallest openings (4 mm, 2 μm, 1 μm, 800 μm, 630 μm, 400 μm, 250 μm, and 125 μm, with a pan at the bottom to collect the dusty fraction). Obtained data were submitted to variance analysis using PROC GLM to evaluate the same effects on feed homogeneity at the release point (beginning, middle and at the end of the feeding alley). The obtained results confirmed what found through Penn State Particle Separator technique, highlighting a decreasing difference in the amount of the rough fraction, collected on the first sieve (4 mm), from 44.12 g, to 40.96 g (DM content). This result could imply, when referred to a TMR with 54% of humidity content, a noticeable difference of 6.75 kg less of rough feed at the end of the feeding alley.

P51. Detectionandidentificationofpoultryprocessedanimalproteins

Van RaamsdonkL.W.D.,BremerM.G.E.G.,PrinsT.W.,Bienemann-PloumM.,ScholtensI.M.J.RIKILT Wageningen UR, Wageningen, The Netherlands

After the Bovine Spongiform Encephalopathy (BSE) crisis emerged in 1985/1986, all processed animal proteins (PAPs) were banned for use in animal feed in the European Union (EU). After development and successful validation of PCR methods to detect PAPs originating from ruminants and pigs, poultry is the last major group which needs proper detection and identification.There are several definitions to describe the term “poultry” in European legislation, partly depending on the purpose and scope. The general definition of “poultry” as intended for safety issues is used as basis. A Dutch project has been started for developing methods for the detection and identification of poultry, based on a consortium of four Dutch companies (SMEs) and RIKILT Wageningen UR. The basic development will be carried out at RIKILT, whereas the SMEs provide materials and take part in the validation studies. The project runs from 2015 until 2018.

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The strategy for developing an integrated approach for the detection and identification of poultry consist of two steps. An immunoassay method will be applied for screening purposes, fit for application in on-site laboratories, and a PCR method will be applied for final identification of positive samples from screening. This step-wise approach implies that the sensitivity of the screening method should be just below the sensitivity of the confirmation method in order to avoid false positives. Furthermore it is not necessary to intend to develop methods with a sensitivity much below the technical requirement of 0.1%, since poultry is considered to be unable to carry and transfer prion diseases.All definitions of poultry consists of species belonging to two different groups (orders) of birds. This situation complicates the use of one specific target. For the PCR method two different targets are used: one for the representatives of the order Galliformes (chicken and turkey) and one for the representatives of the order Anseriformes (duck and geese). The design of the primer-probe combinations allows to use the two sets in one duplex reaction, besides using them independently. The latter situation provides the possibility to distinguish between the two groups for e.g. fraud detection.The chosen approach and design of the methods provides a very versatile way of supporting legislation and enforcement.


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Booth CEN activities

P52. Harmonizationofmethodsofanalysisfornaturaltoxinsinfeed–ActivitiesofCEN/TC327/WG5

Venemans A.1,BreidbachA.2,MulderP.3, Stroka J.2,KujawskiM.4,GrayK.5, Lopez Sanchez P.3,BodiD.6 , Mol H.3

1 Nederlands Normalisatie-instituut (NEN), Delft, The Netherlands 2 European Commission, Directorate General Joint Research Centre, Geel, Belgium3 RIKILT - Wageningen UR, Wageningen, The Netherlands 4 Main Inspectorate of Plant Health and Seed Inspection, Torun, Poland5 LGC, Teddington, UK6 Bundesinstitut für Risikobewertung (BfR), Berlin, Germany BfR

It is important for the European feed sector, national governments and the European Commission that the safety and quality of animal feed, including feed materials, pre-mixtures and feed additives, is guaranteed. When purchasing, producing or selling animal feed, validated and harmonised methods are needed for a uniform judgement of conformity of products to the requirements, in particular in the framework of quality assurance and regulatory control.In the European Union and the European Economic Area, European Standards play an important role in meeting the specific European requirements. The European Committee for Standardization (CEN) is the organisation responsible for European standardization in a wide variety of sectors. One of the technical committees is CEN/TC 327 “Animal feeding stuffs – Methods of sampling and analysis”. CEN/TC 327 drafts methods for organic and inorganic contaminants, mycotoxins, plant toxins, feed additives, veterinary drugs, trace and major elements and parameters related to feed composition. The work of CEN/TC 327 is financially supported by mandates from the European Commission.

Within CEN/TC 327, several working groups (WGs) have been established. WG 5 “Natural toxins” is drafting methods for the analysis of:- T-2, HT-2, DON and ZON by LC-MS/MS (FprEN 16877)- Ergot alkaloids and tropane alkaloids by LC-MS/MS- Mycotoxins (multi-method) by LC-MS/MS- Theobromine by LC-UV and LC-MS/MS- Glucosinolates by LC-MS/MS- Pyrrolizidine alkaloids by LC-MS/MS- Free gossypol by LC-MS/MSFurthermore, a document with criteria for methods of analysis for mycotoxins is under preparation.

One of the most important requirements for standardization is the successful validation of a method by means of a full international collaborative study. The drafting and validation of these methods is in various stages of progress. For some of the work items mentioned, dedicated posters will be presented by the project leaders.

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P53. Harmonizationofmethodsofanalysisforheavymetals,traceelementsandmineralsinfeed–ActivitiesofCEN/TC327/WG4

Venemans A. 1,EykelhoffJ.1,RobouchP.2, Klose R.3, Sloth J.4 , Danier J.5

1 Nederlands Normalisatie-instituut (NEN), Delft, The Netherlands 2 European Commission, Directorate General Joint Research Centre, Geel, Belgium3 Staatliche Betriebsgesellschaft für Umwelt und Landwirtschaft (SMUL), Nossen, Germany4 National Food Institute, Technical University of Denmark (DTU), Söborg, Denmark5 BavarianStateMinistryoftheEnvironmentandConsumerProtection(STMUV),Munich,Germany

It is important for the European feed sector, national governments and the European Commission that the safety and quality of animal feed, including feed materials, pre-mixtures and feed additives, is guaranteed. When purchasing, producing or selling animal feed, validated and harmonised methods are needed for a uniform judgement of conformity of products to the requirements, in particular in the frame¬work of quality assurance and regulatory control.In the European Union and the European Economic Area, European Standards play an important role in meeting the specific European requirements. The European Committee for Standardization (CEN) is the organisation responsible for European standardization in a wide variety of sectors. One of the technical committees is CEN/TC 327 “Animal feeding stuffs – Methods of sampling and analysis”. CEN/TC 327 drafts methods for organic and inorganic contaminants, mycotoxins, plant toxins, feed additives, veterinary drugs, trace and major elements and parameters related to feed composition. The work of CEN/TC 327 is financially supported by mandates from the European Commission.

Within CEN/TC 327, several working groups (WGs) have been established. In WG 4 “Heavy metals, trace elements and minerals” CEN standards have been produced for Dtermination of:- various inorganic substances after pressure digestion by ICP-AES (EN 15621:2012)- selenium by HG-AAS after microwave digestion (EN 16159:2012)- arsenic by HG-AAS after pressure digestion (EN 16206:2012)- mercury by CV-AAS after microwave pressure digestion (EN 16277:2012)- inorganic arsenic by HG-AAS after microwave extraction and separation by SPE (EN

16278:2012)- fluoride content after HAT by ISE (EN 16279:2012)

Currently WG 4 is drafting methods for the analysis of - Trace elements, heavy metals and other inorganic elements in feed by ICP-MS (multi

method) - trace elements, heavy metals and other inorganic elements in feed by ICP-MS- iodine in feed by ICP-MS


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- inorganic arsenic by HPLC-ICP-MS Furthermore, a document with criteria for methods of analysis for heavy metals is

under preparation.

3 CEN standards for the determination of various inorganic substances and heavy metals are under revision:- EN 15510:2007 Determination of calcium, sodium, phosphorus, magnesium,

potassium, iron, zinc, copper, manganese, cobalt, molybdenum and lead by ICP-AES- EN 15550:2007 Determination of cadmium and lead by graphite furnace atomic

absorption spectrometry (GF-AAS) after pressure digestion, and- EN 15621:2012 Determination of calcium, sodium, phosphorus, magnesium,

potassium, sulphur, iron, zinc, copper, manganese and cobalt after pressure digestion by ICP-AES

to enable the proper analysis of lead in kaolinite clay at the regulatory limits set in EU legislation in 2012.


P54. Harmonizationofmethodsofanalysisforadditivesanddrugsinfeed–ActivitiesofCEN/TC327/WG3

Venemans A.1, Mosies C.1,PikkemaatM.2, Genouel C.3, Vincent U.4,ZuidemaT.2,StrnadI.5, PetrováJ.6,vonHolstC.41 Nederlands Normalisatie-instituut (NEN), Delft, The Netherlands 2 RIKILT Wageningen UR, Wageningen, The Netherlands 3 Service Commun des Laboratoires Douanes-Fraudes (SCL), Rennes, France4 European Commission, Directorate General Joint Research Centre, Geel, Belgium5 Österreichische Agentur für Gesundheit und Ernährungsicherheit (AGES), Institut für Tierernährung und Futtermittel, Linz/Vienna, Austria 6 Central Institute for Supervising and Testing in Agriculture (UKZUZ), Prague, Czech Republic

It is important for the European feed sector, national governments and the European Commission that the safety and quality of animal feed, including feed materials, pre-mixtures and feed additives, is guaranteed. When purchasing, producing or selling animal feed, validated and harmonised methods are needed for a uniform judgement of conformity of products to the requirements, in particular in the frame¬work of quality assurance and regulatory control.In the European Union and the European Economic Area, European Standards play an important role in meeting the specific European requirements. The European Committee for

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Standardization (CEN) is the organisation responsible for European standardization in a wide variety of sectors. One of the technical committees is CEN/TC 327 “Animal feeding stuffs – Methods of sampling and analysis”. CEN/TC 327 drafts methods for organic and inorganic contaminants, mycotoxins, plant toxins, feed additives, veterinary drugs, trace and major elements and parameters related to feed composition. The work of CEN/TC 327 is financially supported by mandates from the European Commission.

Within CEN/TC 327, several working groups (WGs) have been established. In WG 3 “Feed additives and drugs” CEN standards have been produced for the isolation and enumeration of various probiotics (EN 1578x-series).

Currently WG 3 is drafting methods for: - Screening on antibiotics by a microbiological plate test- Detection of the antibiotics by Thin Layer Chromatography- Determination of carbadox and olaquindox by HPLC-UV- Identification of antibiotics confirmatory analysis by LC-MS- Determination of organic acids- Determination of authorised coccidiostats- Simultaneous determination of vitamin- Determination of carotenoids- Identification of probiotics


P55. Harmonizationofmethodsofanalysisfororganiccontaminantsinfeed–ActivitiesofCEN/TC327/WG1

Venemans A.1, Hoenstok R.1,vanLeeuwenS.2,FryH.3, Lacoste F.4,deJongJ.21 Nederlands Normalisatie-instituut (NEN), Delft, The Netherlands 2 RIKILT Wageningen UR, Wageningen, The Netherlands 3 Bundesinstitut für Risikobewertung (BfR), Berlin, Germany 4 ITERG, Pessac, France

It is important for the European feed sector, national governments and the European Commission that the safety and quality of animal feed, including feed materials, pre-mixtures and feed additives, is guaranteed. When purchasing, producing or selling animal feed, validated and harmonised methods are needed for a uniform judgement of conformity of products to the requirements, in particular in the framework of quality assurance and regulatory control.


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In the European Union and the European Economic Area, European Standards play an important role in meeting the specific European requirements. The European Committee for Standardization (CEN) is the organisation responsible for European standardization in a wide variety of sectors. One of the technical committees is CEN/TC 327 “Animal feeding stuffs – Methods of sampling and analysis”. CEN/TC 327 drafts methods for organic and inorganic contaminants, mycotoxins, plant toxins, feed additives, veterinary drugs, trace and major elements and parameters related to feed composition. The work of CEN/TC 327 is financially supported by mandates from the European Commission.

Within CEN/TC 327, several working groups (WGs) have been established. In WG 1 “Organic contaminants” CEN standards have been produced for:- Dioxins and dioxin-like PCBs by GC/HRMS and of non-dioxin-like PCBs by GC/HRMS

(EN 16215:2012)- Organochlorine pesticides and non-dioxin-like PCBs by GC/MS (EN 15741:2009)- Organochlorine pesticides and non-dioxin-like PCBs by GC/ECD (EN 15742:2009)

Currently WG 1 is drafting methods for the analysis of:- Melamine and cyanuric acid by LC-MS/MS- Mineral oil by GC-FID- Pentachlorophenol by LC-MS

Furthermore the abovementioned CEN standards for organochlorine pesticides and non-dioxin-like PCBs will be revised: EN 15741:2009 to enable the determination of PCBs at the regulatory limits set in EU

legislation in 2012 EN 15741:2009: the NDL-PCBs will be deleted from the scope because the required

sensitivity cannot be reached with ECD; 4 additional organochlorine pesticides will be added to the scope.


P56. Harmonizationofmethodsofsamplingandanalysisoffeed–ActivitiesofCEN/TC327

Veneman A.1, Mosies C. 1, Brenten T.2,AltzitzoglouT.3,EllermannN.4,deJongJ.51 Nederlands Normalisatie-instituut (NEN), Delft, The Netherlands 2 Mars Pet Care, Verden, Germany, on behalf of European Pet Food Industry Association (FEDIAF)3 European Commission, Directorate General Joint Research Centre, Geel, Belgium

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4 Ministeriet for Fødevarer, Landbrug og Fiskeri, Ringsted, Denmark5 RIKILT Wageningen UR, Wageningen, The Netherlands

It is important for the European feed sector, national governments and the European Commission that the safety and quality of animal feed, including feed materials, pre-mixtures and feed additives, is guaranteed. When purchasing, producing or selling animal feed, validated and harmonised methods are needed for a uniform judgement of conformity of products to the requirements, in particular in the framework of quality assurance and regulatory control.In the European Union and the European Economic Area, European Standards play an important role in meeting the specific European requirements. The European Committee for Standardization (CEN) is the organisation responsible for European standardization in a wide variety of sectors. One of the technical committees is CEN/TC 327 “Animal feeding stuffs – Methods of sampling and analysis”. CEN/TC 327 drafts methods for organic and inorganic contaminants, mycotoxins, plant toxins, feed additives, veterinary drugs, trace and major elements and parameters related to feed composition. The work of CEN/TC 327 is financially supported by mandates from the European Commission.

Within CEN/TC 327, several working groups (WGs) have been established. The WGs on Composition (WG 1), Feed additives and drugs( WG 3), Heavy metals, trace elements and minerals (WG 4) and Natural toxins (WG 5) are covered separately.

WG 2 “Composition” has developed a standard for - The determination of acid detergent fibre (ADF) and acid detergent lignin (ADL)

contents (EN ISO 13906:2008)and is currently drafting a method for- The determination of energy value in pet food (prEN 16967)Furthermore, WG 2 supports the relevant ISO group with the revision of - EN ISO 12099: 2010 Guidelines for the application of near infrared spectrometry- CEN ISO TS 17764:2006 (2 parts) on the determination of fatty acids.

WG 6 “Radioactivity measurements” will develop a method for - The determination of the radionuclides 131I,134Cs and 137Cs in feed.


CEN/TC 327 is also cooperating with ISO/TC 34/SC 10 “Animal feeding stuffs” on the parallel development of European and global standards (EN ISOs).


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Delegates

Delegate Institute Acar Ümit Mugla Sitki Kocman UniversityAchten Elisabeth Federal Institute for Risk Assessment (BfR)Adamse Paulien Wageningen University & ResearchAmato Giuseppina Istituto Zooprofilattico SperimentaleAmlund Heidi National Institute of Nutrition and Seafood (NIFES)Antonissen yara EU Association of Specialty Feed Ingredients and their Mixtures (FEFANA)Arnaud Ludovic Lallemand sasBaeten Vincent Walloon Agricultural Research Centre (CRA-W)Bellorini Stefano European Commission - JRC GeelBergsma Nynke DuPointBerntssen Marc National Institute of Nutrition and Seafood (NIFES)Bodi Dorina Federal Institute for Risk Assessment (BfR)Boix Ana European Commission - JRC GeelBraeckman Hans Primoris BelgiumBucci Giulio EU Association of Specialty Feed Ingredients and their Mixtures (FEFANA)Brereton Paul FERACapodieci Giuseppe Luca EU Association of Specialty Feed Ingredients and their Mixtures (FEFANA) Chedin Mostafa European Commission -JRC GeelCodony Rafael University of BarcelonaDardenne Pierre Walloon Agricultural Research Centre (CRA-W)Dejaegher Yvan Belgian Compound Feed Industry Associationde Jong Jacob RIKILT Wageningen UREisenberg David Micro-Tracers,Inc.Eisenberg Zachary Micro-Tracers,Inc.Ezerskis Zigmas European Commission -JRC GeelFernández Juan Antonio Walloon Agricultural Research Centre (CRA-W)Fournier Annie Canadian Food Inspection AgencyFry Hildburg Federal Institute for Risk Assessment (BfR)Fumiere Olivier Centre wallon de Recherches agronomiques (CRA-W)Geinoz Michel Federal Department of Economic Affairs, Education and Research EAER AgroscopeGenouel christophe Service Commun des Laboratoires DGCCRF et DGDDI

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Gerritsen Henk RIKILT Wageningen URGonzalez de la Huebra Maria Jose European Commission -JRC GeelGort Frank SecureFeedGrandič Marjana University od Ljubljana, Veterinary facultyHan Lujia China Agricultural UniversityHelland Ellinor BioMar ASHoogenboom Ron RIKILT Wageningen URHumbert Marc Eurofins Analytics FranceKarus Avo Estonian university of life sciencesKesbic Osman Sabri Kastamonu UniversityKruse Sabine Federal Ministry of Food and AgricultureLavrijsen Lucas LV LUC LAVRIJSENLiu Xian China Agricultural UniversityMarounek Milan Institute of Animal ScienceMcCarron Eileen State LaboratoryMoll Wulf-Dieter BIOMIN Holding GmbHNordkvist Erik National Veterinary Institute, SVANumata Jorge Federal Institute for Risk Assessment (BfR), GermanyOernsrud Robin National Institute of Nutrition and Seafood Research Omar Jone European Commission -JRC GeelOttoboni Matteo University of MilanPinotti Luciano University of MilanPonghellini Marta EU Commission DG Health and Food SafetyQuillama Torres Liliana GMP+ InternationalRadewahn Peter Deutscher Verband Tiernahrung e. V.Rasinger Josef Daniel National Institute of Nutrition and Seafood ResearchRobert Christelle CER GroupeRobouch Piotr European Commission -JRC Geel Sanden Monica National Institute of Nutrition and Seafood (NIFES)Seifert Jörg EU Association of Specialty Feed Ingredients and their Mixtures (FEFANA)Serano Federica European Commission -JRC GeelSibanda Liberty Randox Food DiagnosticsSloth Jens National Food Institute (DTU Food)Slowikowski Boleslaw European Commission -JRC GeelTavcar-Kalcher University of Ljubljana, Veterinary FacultyTen Dam Guillaume RIKILT Wageningen URThelander Magnus Swedish National Veterinary InstituteTohill Aaron Randox Food DiagnosticsTretola Marco University of MilanUrdl Marcus National Veterinary Institute Sweden, SVAVan der Boon Els GMP+ Internationalvan Raamsdonk Leo Wageningen University & Research


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van Vuure Carine Darling Ingredients International Holding BVVancutsem Jeroen (ir.) Federal Agency for the Safety of the Food ChainVandaele Bill BILL SUPPORTVandicke Jonas University of GhentVenemans Annemieke CEN/TC 327Vermeulen Philippe Walloon Agricultural Research Centre (CRA-W)Verstraete Frans European Commission, DG Health and Food SafetyVincent Ursula European Commission -JRC Geelvon Holst Christoph European Commission -JRC GeelVukmiroviƒá Đuro University of Novi Sad, Institute of food technologyWegh Robin RIKILT Wageningen URYang Zengling China Agricultural University

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19 - 20 October 2016 Geel - Belgium


Programme & Abstract book


Feed 20

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feed 2016feedsafety.org/wp-content/uploads/2014/02/programme-book2016.pdf · 2 general introduction...

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