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Dr. Rafael Codony

Dr. Rafael Codony. 6th Framework Programme of Research - EC. “ Quality and safety of feeding fats obtained from co-products or by-products from the food chain ” (Coordinator: Dr. Rafael Codony) http://www.ub.edu/feedfat/. rafaelcodony@ub.edu. PARTNERS of the project:

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Dr. Rafael Codony

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  1. Dr. Rafael Codony 6th Framework Programme of Research - EC “Quality and safety of feeding fats obtained from co-products or by-products from the food chain” (Coordinator: Dr. Rafael Codony) http://www.ub.edu/feedfat/ rafaelcodony@ub.edu

  2. PARTNERS of the project: 1-Dept of Food Science & Nutrition, University of Barcelona 2-Dept of Food Science,Swedish University of Agric. Science, Uppsala 3-Department of Food Science, University of Bologna 4-Lab. Physico & Toxico Chimie, University of Bordeaux 1-CNRS -LPTC 5-Ecotecnologies Department,IIQAB-CSIC, Barcelona 6-Animal and Food Science Department, Universitat Autònoma de Barcelona 7-Animal Science Department, Universitat Politècnica de Valencia 8-Stazione Sperimentale Oli e Grasse, Milano 9-SILO s.r.l., Firenze

  3. Objectives of the project were designed trying to answer the following questions: • What are the main chemical characteristics of these fats? • Are fat by- or co-products really safe for animal production? • How contribute these fats to the global intake of contaminants (dioxins, PCB, PBDE, PAH) through the diet? • Which are the repercussions of using these fats on lipid quality and stability of meat? • What are the suitable analytical controls to assess quality and safety of fats? How standardisation and policy control of these fats could be improved? • Are consumers confident respect to these systems of production ? • Are these fats really available, useful and profitable for fat and feed producers?

  4. PART I: FAT CHARACTERIZATION “Quality and safety of feeding fats obtained from co-products or by-products from the food chain”

  5. Moisture Fatty acid composition Total Cholesterol Content Sterol composition Tocopherols/Tocotrienols Trans Fatty acids Conjugated Fatty Acids Acid Value Mono- and diacylglycerols FTIR spectra Sterol oxidation products Fatty acid oxidation products Polymer content Peroxide Value (volumetric) Peroxides (XO colorimetric) TBA value p-anisidine value Dioxins and PCBs PAHs PBDEs Fats collected directly from factories in 10 European countries: 1.Acid oils from chemical refining (n=25)AOCHE 2.Acid oils from physical refining (n=16)AOPHY 3.Lecithins (n=8)LECI 4.Recycled cooking oils(n=8)RECY 5.Animal fats (beef tallow, lard, mixed origin) (n=36)ANFA 6.Oils extracted from exhausted bleaching earths (n=2)EBE 7.Fish oils (n=9)FISH 8.Hydrogenated fats from by/co-products (n=6)HYBY 9.Fatty acid calcium soaps (n=3)FACS

  6. FT-IR spectroscopy for classification of fats Classification of feeding fats by FT-IR spectroscopy. G. Gasperini et al. (2007) Eur. J. Lipid Sci. Technol. 109 (2007) 673–681 FTIR spectra

  7. FT-IR spectroscopy for classification of fats Classification tree

  8. FT-IR spectroscopy for classification of fats • Only 8 samples out of 120 (94.6%) were not correctly assigned to their category. ANFA, RECY ans EBA are not well separated • Almost all these 8 samples also showed anomalous chemical values (i.e. acidity) • To complete la clasification values or another parameters could be used (i.e. % polymers (always >4% for EBE y RECY and <4% for ANFA • It must be validated with a higher number of samples, for certain categories of fat materials • Problems: (1) Lack of standardisation of fat materials; (2) Presence of blends

  9. Lipid Composition Values C.Nuchi, F.Guardiola, R.Bou, P.Bondioli, L.Della Bella, R.Codony. J Agric Food Chem (2009) 57, 1952-1959 * Results expressed as: min – max (median; average) ; ** Compensated area normalization

  10. Lipid Degradation Values C.Nuchi, F.Guardiola, R.Bou, P.Bondioli, L.Della Bella, R.Codony. J Agric Food Chem (2009) 57, 1952-1959 Parameters related to lipid oxidation * Results expressed as: min – max (median; average); ** NA: not analyzed (in some cases, due to solubility problems)

  11. Lipid Degradation Values • Peroxide Value is not discriminant. • Secondary oxidation is quite variable between categories, showing Acid Oils, Lecithins and Fish oils the highest values. • Polymers are very relevant in EBE and RECY, and intermediate in FISH oils. • (!) Problem: Lack of QUALITY standards

  12. Moisture and Acid Value * Results expressed as: min – max (median; average); NA: samples not analyzed

  13. Acid Value is highly variable between categories, but also inside some of them • (!) Problem: Lack of QUALITY standards

  14. RESULTS: Persistentcontaminants in fatmaterials “Quality and safety of feeding fats obtained from co-products or by-products from the food chain”

  15. Dioxins (PCDDs/PCDF) and PCBs in fats PCDD/Fs and DL-PCBs in feeding fats obtained as co-products or by-products derived from the food chain. Ábalos, M. et al. Chemosphere 71 (2008) 1115–1126 FISH oils PCDD/Fs + DL-PCBs maximum level !!! • FISH OIL samples showed the highestPCDD/F and DL-PCB concentrations. This fact was expected due to the animal origin of these products. But only 1 sample exceededthe maximum levels established for these pollutants at the Commission Directive 2006/13/EC (6 pg WHO-TEQ of PCDD/F and 24 pg WHO-TEQ of PCDD/F+DL-PCB/ g oil).

  16. PCDDs/PCDFs and PCBs in fats • All the analyzedanimal fatsshowed levels of PCDD/Fs and DL-PCBs below the maximum establishedat the Commission Directive. • Samples of vegetal origin(i.e. LECI, HYBY, AOCHE) showed, in general,very low concentrations of PCDD/Fs and DL-PCBs. 1.25 ng WHO-TEQ/ kg 3.0 ng WHO-TEQ/kg (animal fats) 24 ng WHO-TEQ/kg (fishoils) (feedmaterials of plant origin) 1.5 ng WHO-TEQ/ kg (vegetable oils and by-products) (PCDDs/Fs + DL-PCBs), expressed in pg WHO-TEQ/g) • Special attention should be paid toAOPHYsince seven of a total of nine samplesexceeded the maximum level for PCDD/Fset at the Commission Directive for by-products of vegetable oils. • In addition,five of these seven samplesalso exceeded the maximum set for the sum of PCDD/Fs and DL-PCBs.

  17. Total [PAHs] (ng/g) : min - max (median) levels Samples with Fat LMW-PAHs MMW-PAHs HMW-PAHs Total PAHs [BaP]>2 ng/g ANFA 1 - 25 (2) 1 - 90 (15) 1 - 54 (1) 3/36 <10 - 107 (18) AOCHE 1 - 1 540 (304) 16 - 4 040 (1043) 1 - 340 (20) 15/25 21 - 4989 (1022) AOPHY 10 - 5 990 (193) 60 - 24 520 (864) 1 - 190 (25) 11/16 51 - 30700 (521) EBE 149 - 155 (152) 447 - 466 (457) 8 - 56 (32) 2/2 610 - 672 (641) FACS 25 - 102 (34) 308 - 447 (355) 14 - 61 (16) 0/3 356 - 610 (396) FISH 7 - 43 (20) 10 -160 (33) 1 - 9 (3) 0/9 <10 - 177 (61) HYBY 15 - 38 (23) 7 - 280 (230) 3 - 24 (7) 1/6 56 - 318 (269) LECI 1 - 26 (10) 5 - 140 (20) 1 - 8 (4) 0/9 <10 - 155 (31) MIX 1 - 240 (17) 8 - 1 080 (18) 1 - 62 (8) 1/10 19 - 1394 (65) RECY 11 - 136 (25) 2 - 243 (37) 1 - 103 (7) 2/8 28 - 482 (61) Polycyclic Aromatic Hydrocarbons (PAHs) in fats PAHs: First EU regulation in 2006 2 ng/g for BaP (highest carcinogenic potential) in oils and fats intended for direct human consumption or use as an ingredient in food (Com. Reg. EC No 1881/2006). NO regulation for meat or for feedstuffs for animal feeding Low molecular weightPAHs (LMW) : N, Ace, Acy, Fe; Medium molecular weightPAHs (MMW) : Phe, A, Fluo, Pyr, BaA, Triph + Chrys; High molecular weightPAHs (HMW) : BbF + BkF, BaP, BeP, Per, IP, DahA + DacA, BP.

  18. Polycyclic Aromatic Hydrocarbons (PAHs) in fats • 3 levels of contamination(related to levels of BaP): • - very low (<0.5 – 2 ng/g) : FISH, HYBY, LECI • - very low but potentially at risk : ANFA, EBE, FACS, RECY • - low to very high : AOCHE, AOPHY 36 samples Animal Fats

  19. PART II: ANIMAL TRIALS “Quality and safety of feeding fats obtained from co-products or by-products from the food chain”

  20. HIGH Trans FA MEDIUM Trans FA LOW Trans FA HIGH PAHs MEDIUM PAHs LOW PAHs HIGH Oxidated MEDIUM Oxidated LOW Oxidated HIGH Diox-PCBs MEDIUM Diox-PCBs LOW Diox-PCBs Selection of Fat Materials and Feed Preparation Effects on animals and meat TRIAL 1 TRIAL 3 TRIAL 4 TRIAL 2 • Lipid composition • Contents of trans and CLA isomers • Lipid degradation levels • Contaminants: contents of dioxines and PCBs, PAHs and their metabolites, and PBDEs • Levels of contaminants in meat, and other tissues f) Productive parameters: Daily feed intake, Liveweight gain, Feed efficiency, Carcass yield, and Meat yield g) Digestibility trials h) Animal health parameters: -Histology of intestinal mucosa (villus height/crypt depht ratio) -Cellular composition in gut associated lymphoid tissues -Characterization of the microbial ecosystem in hindgut (T-RFLP) -Hindgut contents of short chain fatty acids (by GLC)

  21. TRIAL 5 Transfer of contaminants Diet 1 Background Diet 2 ½ Max level Diet 3 *Max Level Diet 4 2 Max level PAHs: Phe+Fluo+Chrys+BeP+BaP (1) not addition (2) 300 ng/g feed (3) 600 ng/g feed (4) 1200 ng/g feed PCDD/PCDFs: 17 toxic congeners (1) not addition (2) 0.38 pg WHO-TEQ/g feed (3) 0.75 pg/g feed (4) 1.50 pg/g feed DL-PCBs: 12 toxic congeners (1) not addition (2) 0.38 pg WHO-TEQ/g feed (3) 0.75 pg/g feed (4) 1.50 pg/g feed PBDEs: BDE 47+BDE 153 (1) not addition (2) 3 ng/g feed (3) 6 ng/g feed (4) 12 ng/g feed • PAHs determination (GC-MS) in oils, feeds, meats & livers • OH-PAHs determination (UPLC-MS/MS) in plasma, bile & excreta • PBDEs determination (GC-ECD) in oils, feeds, meats & livers • PCDD/PCDFs determination (HRGC-HRMS) in oils, feeds, meats & livers • DL-PCBs determination (HRGC-HRMS) in oils, feeds, meats & livers *MAX levels were established according to EC regulations, or to max levels found in 1st Part

  22. Conclusions about the effects on animal production and health • In summary, under our experimental conditions and based on all the compiled data, we can conclude that the inclusion in chicken or rabbit feed of different fat co- and by-products differing in levels of trans fatty acids, dioxins and PCBs, PAHs, or lipid oxidation had not major effect on animal health. Some of the differences found between treatments are more dependant on the origin, quality and profile of the dietary fat, than on the alteration or contamination itself. • Some negative repercussions on meat quality and animal performance were only observed for rabbits in one treatment with fish oil (but it was the treatment LOW in dioxin-PCB content). Similar problems were not oberserved for chickens with the same treatment. This is another input indicating that low quality fish oils can be easily found, and reveal the relevance of their control of production and traceability.

  23. 4 Experimental Feeds DP-1:Background DP-2:½ max: 0.38 pg WHO-TEQ/g (PCDD/Fs) and 0.38 pg WHO-TEQ/g (DL-PCBs) DP-3: max:0.75 pg WHO-TEQ/g (PCDD/Fs) and 0.75 pg WHO-TEQ/g (DL-PCBs) DP-4:2 max: 1.5 pg WHO-TEQ/g (PCDD/Fs) and 1.5 pg WHO-TEQ/g (DL-PCBs) Meat (chicken with skin) Maximum levels for meat and meat products of poultry and farmed game (COMMISSION REGULATION (EC) Nº 199/2006 3 February 2006): 2 pg WHO-TEQ/g fat PCDD/Fs 4 pg WHO-TEQ/g fat PCDD/Fs + DL-PCBs !! Levels exceeding MAX levels regulated in meats !! Results on transfer of Dioxins/PCBs from feeds to meat Meat DP-1 Meat DP-2 Meat DP-3 Meat DP-4 pg WHO-TEQ/g fat PCDD/Fs 0.21 3.16 6.50 12.57 Total (PCDD/Fs + DL-PCBs) 0.29 6.79 13.80 27.02

  24. Conclusions on transfer of Dioxins/PCBs • Chicken meat samples (including skin)exceed the maximum PCDD/F and DL-PCB levels established at the EC Regulation, although corresponding levels in feed were under max levels. • Remarkable lower levels of concentrations of PCDD/Fs were found in rabbit compared to chicken meat, never exceeding maximum levels regulated. • Good correlation between congener distribution profiles in the feeds and in the meat samples were found for chickens, but not for rabbits. • The lower rate of transfer to tissues observed in the rabbit could be due to 3 main factors: - 3% fat was added to rabbit feed meanwhile 6 % to chicken feed - Fat in rabbit meat (3%) is much lower than in chicken meat+skin (12%) - Bioaccumulation of dioxins/PCBs in the rabbit follows a different pattern than in the chicken

  25. Oils and feeds SPIKE trial(Phe, Fluo, Chrys, BeP, BaP) PAH trial(high level) [PAH] ng/g Total PAHs BaP Carcinogenic [PAH] ng/g Total PAHs BaP Carcinogenic Oil (AOCHE) 5300 90 800 (15%) 1/2 x max 900 200 400 (40%) [PAH] ng/g Total PAHs BaP Carcinogenic Max 1900 400 800 (40%) Rabbit feed 150 2.5 25 (17%) 2 x max 3900 800 1600 (40%) * carcinogenic : BaA & BaP Chicken feed 300 6 50 (17%) Animal tissues • No transfer to animal meat, even when using highly contaminated oils from chemical or physical refining, or with highest spiked levels. • No effect ofhigh or extremelyhigh PAH contents in feeds ongrowth and health performances ofanimals • At he highest levels, detectable amounts of some PAHs metabolites are found in plasma Results on transfer of PAHs from feeds to meat

  26. Results & Conclusions on transfer of PAHs/PBDEs • PAHs present in the added fat were persistent in the final feed (70-90% recovery). • No risk for animal health and for the consumer of the meat, even with very high levels of PAHs in feed fats. • Use of PAH metabolites in plasma as good biomarkers of PAH exposure, but only when very high levels were present in the feed fat. • In consequence, at the highest levels of PAH contamination it can exist certain risk for animal health, since they can be distributed from plasma to tissues. • Determination of levels of these metabolites in meat should be made in order to know if it could exist some risk for the consumer. • PBDEs showed a high rate of bioaccumulation, in proportion to the feed concentration. But low risk can be expected since PBDE values found in fats were too small.

  27. Other studies included in the project • Proposal of analytical control programs for policy purposes • (Docs “Analytical control program” and “Handbook of analytical methods” can be found at http://www.ub.edu/feedfat/documents.htm , page documents, Maindocuments) • Socio-economic impact of the application of fatty co- and by-products in commercial feeds • -Parini, M., Cantini, F. (Parte I) Riv. Ital. SostanzeGrasse 2008, 85(4), 246-265. • -Parini, M., Cantini, F. (Parte II)" Riv. Ital. SostanzeGrasse 2009, 86(1), 1-72. • Consumer acceptance study • (CONSUMER OPINION can be found at http://www.ub.edu/feedfat/documents.htm , page documents, Final meetingpresentations)

  28. Global conclusions “Quality and safety of feeding fats obtained from co-products or by-products from the food chain”

  29. They are some fat materials obtained as co- and by-products of the food chain that can provide valuable characteristics in feeding animals (i.e. animal fats and acid oils). But in some cases, a higher level of standardization should be promoted. • For some fat categories (i.e. AOPHY), adequate system of purification should be studied in order to increase quality and safety of their use. Other fats need only a better system of traceability control and good manufacturing practices to reach suitable quality levels. • PAHs do not accumulate in animal tissues and have scarce repercussion on safety of foods from animal origin. In any case, extra-costs for the supplementary filtration with active carbon are negligible and acceptable from economical point of view.

  30. In contrast, dioxins and DL-PCBs are highly accumulated in animal tissues because of their liposolubility. This fact pose a question about the non reliable relationship between the regulated feed maximum levels of dioxins and DL-PCBs and the maximum levels regulated in meat and other foods from animal origin. • From the comparison of costs of iso-energetic feed formulas containing co-products or conventional oils, we can conclude that the use of co-products such as acid oils and animal fats are economically interesting alternatives for feed and meat producers if compared with conventional oils such as soybean, rape, sunflower and palm oil. • From our analysis resulted that the advantages of the use of certain co-products and technical lipids in feed are independent from environmental benefits. Their use has reasons in itself and not as an alternative use to disposal.

  31. Dr. Rafael Codony 6th Framework Programme of Research - EC “Quality and safety of feeding fats obtained from co-products or by-products from the food chain” Detailed information about results & conclusions on the different aspects studied can be found in reported articles, and at: http://www.ub.edu/feedfat/documents.htm rafaelcodony@ub.edu

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