SCIENTIFIC OPINION. EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) 2,3

Transcription

1 SCIENTIFIC OPINION Scientific Opinion on the safety and efficacy of copper compounds (E4) as feed additives for all species: cupric chelate of amino acids hydrate, based on a dossier submitted by Zinpro Animal Nutrition Inc. 1 EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) 2,3 European Food Safety Authority (EFSA), Parma, Italy This scientific output, published on 10 January 2014, replaces the earlier version published on 21 February ABSTRACT Cupric chelate of amino acids hydrate is safe for all animal species/categories up to the authorised maximum of total copper content in complete feed. Consumption surveys include copper from foodstuffs of animal origin. Since the supplementation of animal feed with copper-containing compounds has not essentially changed over the last decade, no change in the contribution of foodstuffs originating from supplemented animals to the overall copper intake of consumers is expected. No concerns for consumer safety are expected from the use of cupric chelate of amino acids hydrate in animal nutrition, which would substitute for other copper sources. The additive should be considered as a skin and eye irritant and, owing to its amino acid/peptide component, as a skin/respiratory sensitiser. Potential risks to soil organisms have been identified as a result of the application of piglet manure. Levels of copper in other types of manure are too low to create a potential risk within the timescale considered. There might also be a potential environmental concern related to the contamination of sediment resulting from drainage and the run-off of copper to surface water. In order to draw a final conclusion, further model validation is needed and some further refinement to the assessment of copper-based feed additives in livestock needs to be considered, for which additional data would be required. The use of copper-containing additives in aquaculture up to the authorised maximum of total copper content in complete feeds is not expected to pose an appreciable risk to the environment. The extent to which copper-resistant bacteria contribute to the overall antibiotic resistance situation cannot be quantified at present. Cupric chelate of amino acids hydrate is recognised as an efficacious source of copper to meet animal requirements. European Food Safety Authority, 2013 KEY WORDS Nutritional additive, compounds of trace elements, cupric chelate of amino acids hydrate, safety, environment, efficacy 1 On request from the European Commission, Question No EFSA-Q , adopted on 31 January Panel members: Gabriele Aquilina, Alex Bach, Vasileios Bampidis, Maria De Lourdes Bastos, Gerhard Flachowsky, Josep Gasa-Gasó, Mikolaj Antoni Gralak, Christer Hogstrand, Lubomir Leng, Secundino López-Puente, Giovanna Martelli, Baltasar Mayo, Derek Renshaw, Guido Rychen, Maria Saarela, Kristen Sejrsen, Patrick Van Beelen, Robert John Wallace and Johannes Westendorf. Correspondence: FEEDAP@efsa.europa.eu 3 Acknowledgement: The Panel wishes to thank the members of the Working Group on Trace Elements, including Noël Albert Dierick, Jürgen Gropp, Alberto Mantovani, Joop de Knecht and the late Reinhard Kroker, for the preparatory work on this scientific opinion. 4 Revision 1: erratum. The recommendation concerning the molecular weight of the compound has been redefined. Suggested citation: EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP); Scientific Opinion on the safety and efficacy of copper compounds (E4) as feed additives for all species: cupric chelate of amino acids hydrate, based on a dossier submitted by Zinpro Animal Nutrition Inc... [26 pp.]. doi: /j.efsa Available online: European Food Safety Authority, 2013

2 SUMMARY Following a request from the European Commission, the Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) was asked to deliver a scientific opinion on the safety and efficacy of cupric chelate of amino acids hydrate when used as feed additive for all animal species. Cupric chelate of amino acids hydrate is safe for all animal species/categories up to the authorised maximum of total copper content in complete feed. Consumption surveys include copper from foodstuffs of animal origin. Since the supplementation of animal feed with copper-containing compounds has not essentially changed over the last decade, no change in the contribution of foodstuffs originating from supplemented animals to the overall copper intake of consumers is expected. No concerns for consumer safety are expected from the use of cupric chelate of amino acids hydrate in animal nutrition, which would substitute for other copper sources. The additive should be considered as a skin and eye irritant and, owing to its amino acid/peptide component, as a skin/respiratory sensitiser. Potential risks to soil organisms have been identified as a result of the application of piglet manure. Levels of copper in other types of manure are too low to create a potential risk within the timescale considered. There might also be a potential environmental concern related to the contamination of sediment resulting from drainage and the run-off of copper to surface water. In order to draw a final conclusion, further model validation is needed and some further refinement to the assessment of copper-based feed additives in livestock needs to be considered, for which additional data would be required. The use of copper-containing additives in aquaculture up to the authorised maximum of total copper content in complete feeds is not expected to pose an appreciable risk to the environment. The extent to which copper-resistant bacteria contribute to the overall antibiotic resistance situation cannot be quantified at present. Cupric chelate of amino acids hydrate is recognised as an efficacious source of copper to meet animal requirements. 2

3 TABLE OF CONTENTS Abstract... 1 Summary... 2 Table of contents... 3 Background... 4 Terms of reference... 5 Assessment Introduction Identity and characterisation Manufacturing process Characterisation of the copper compound Characterisation of the additive Physical state of the additive Stability Homogeneity Physico-chemical incompatibilities in feed Conditions of use Evaluation of the analytical methods by the European Union Reference Laboratory (EURL) Safety Safety for the target species Tolerance of target animals Microbial studies Conclusions on safety for the target species Safety for the consumer Metabolic and residue studies Assessment of consumer safety Conclusions on the safety for the consumer Safety for the users/workers Safety for the environment Conclusions on safety for the environment Efficacy Post-market monitoring Conclusions and recommendations General remark Documentation provided to EFSA References Appendices

4 BACKGROUND Regulation (EC) No 1831/ establishes the rules governing the Community authorisation of additives for use in animal nutrition. Article 10(2) of that Regulation also specifies that for existing products within the meaning of Article 10(1), an application shall be submitted in accordance with Article 7, at the latest one year before the expiry date of the authorisation given pursuant to Directive 70/524/EEC for additives with a limited authorisation period, and within a maximum of seven years after the entry into force of this Regulation for additives authorised without time limit or pursuant to Directive 82/471/EEC. The European Commission received a request from the company Zinpro Animal Nutrition Inc. 6 for reevaluation of authorisation of the copper-containing additive cupric chelate of amino acids hydrate (Availa Cu) when used as a feed additive for all animal species (category: Nutritional additives; functional group: compounds of trace elements). According to Article 7(1) of Regulation (EC) No 1831/2003, the Commission forwarded the application to the European Food Safety Authority (EFSA) under Article 10(2) (re-evaluation of an authorised feed additive). EFSA received directly from the applicants the technical dossier in support of this application. 7 According to Article 8 of that Regulation, EFSA, after verifying the particulars and documents submitted by the applicant, shall undertake an assessment in order to determine whether the feed additive complies with the conditions laid down in Article 5. The particulars and documents in support of the application were considered valid by EFSA as of 21 September The additive cupric chelate of amino acids hydrate had been authorised in the EU under the element Copper-Cu for all animal species Without a time limit (Commission Regulation (EC) No 1334/2003) 8 and amendments. Following the provisions of Article 10(1) of Regulation (EC) No 1831/2003 the compound was included in the EU Register of Feed Additives under the category Nutritional additives and the functional group Compounds of trace elements. 9 The Scientific Committee on Animal Nutrition (SCAN) delivered reports on the use of copper methionate for pigs (EC, 1981), copper compounds in feedingstuffs (EC, 1982) and in feedingstuffs for pigs (EC, 1983) and the use of copper in feedingstuffs (EC, 2003a). 10 EFSA issued opinions on the safety of the chelated forms of iron, copper, manganese and zinc with synthetic feed grade glycine (EFSA, 2005), on the safety and efficacy of a copper chelate of hydroxy analogue of methionine (Mintrex Cu) as feed additive for all species (EFSA, 2008a; EFSA, 2009a), and on the safety and efficacy of di copper chloride tri hydroxide (tribasic copper chloride, TBCC) as feed additive for all species (EFSA, 2011). EFSA has issued an opinion concerning the re-evaluation of cupric sulphate pentahydrate (EFSA, 2012). 5 Regulation (EC) No 1831/2003 of the European Parliament and of the Council of 22 September 2003 on additives for use in animal nutrition. OJ L 268, , p Zinpro Animal Nutrition Inc.. Gerard Doustraat 4a Boxmeer. The Netherlands. 7 EFSA Dossier reference: FAD Commission Regulation (EC) No 1334/2003 of 25 July 2003 amending the conditions for authorisation of a number of additives in feedingstuffs belonging to the group of trace elements. OJ L 187, , p European Union Register of Feed Additives pursuant to Regulation (EC) No 1831/2003. Available from 10 Available from 4

5 TERMS OF REFERENCE According to Article 8 of Regulation (EC) No 1831/2003, EFSA shall determine whether the feed additive complies with the conditions laid down in Article 5. EFSA shall deliver an opinion on the safety for the target animals, consumer, user and the environment and the efficacy of the Cupric chelate of amino acids hydrate, when used under the conditions described in Table 1. 5

6 Table 1: Description and conditions of use of the additive as proposed by the applicant Zinpro Animal Nutrition Inc. Additive Registration number/ec No/No (if appropriate) Category(-ies) of additive Functional group(s) of additive Availa Cu Nutritional additive 3(b) Composition, description Copper Amino Acid Chelate, Hydrate Chemical formula Description Cu (x) 1-3 nh 2 O (x = anion of any amino acid derived from hydrolysed soya protein) Molecular weight not exceeding Purity criteria Method of analysis (if appropriate) (if appropriate) 10% Copper Copper assay using CEN/TS 15621: Animal feeding stuffs- Determination of calcium, sodium, phosphorus, magnesium, potassium, sulphur, iron, zinc, copper, manganese, cobalt and molybdenum after pressure digestion by ICP-AES. Trade name (if appropriate) Name of the holder of authorisation (if appropriate) Availa Cu Zinpro Animal Nutrition Inc., The Netherlands Conditions of use Species category animal All species or of Maximum Age See specifics by species Maximum content mg/kg of complete feedingstuffs Pigs - piglets up to 12 weeks: - other pigs: Bovine 1. Bovine before the start of rumination: - milk replacers - other complete feedingstuffs 2. Other bovine: Ovine Fish Crustaceans Other species 170 (total) 25 (total) 15 (total) 15 (total) 35 (total) 15 (total) 25 (total) 50 (total) 25 (total) Withdrawal period (if appropriate) Not relevant 6

7 Other provisions and additional requirements for the labelling Specific conditions or restrictions for use (if appropriate) Specific conditions or restrictions for handling (if appropriate) Post-market monitoring (if appropriate) Specific conditions for use in complementary feedingstuffs (if appropriate) The following declarations shall be inserted in the labeling and accompanying documents: For sheep: Where the level of copper in feedingstuffs exceeds 10 mg/kg: the level of copper in this feedingstuff may cause poisoning in certain breeds of sheep. For bovines after the start of rumination: Where the level of copper in feedingstuffs is less than 20 mg/kg: the level of copper in this feedingstuff may cause copper deficiencies in cattle grazing pastures with high contents of molybdenum or sulphur. For user safety: gloves, respiratory and eye protection recommended. (see MSDS) Tracking & tracing will be in full place, all remarks during the use of Availa Cu will be noticed and recorded. Availa Cu can be mixed in feeds, to supply Cu in final feed within EU legal limits for each species. Maximum Residue Limit (MRL) (if appropriate) Species or category of Target tissue(s) or food Maximum content in Marker residue animal products tissues Non applicable Non applicable Non applicable Non applicable 7

8 ASSESSMENT This opinion is based in part on data provided by an applicant involved in the production/ distribution of copper-containing compounds. It should be recognised that these data covers only a fraction of the existing cupric chelate of amino acids hydrate. 1. Introduction The biological role of copper, its requirements/recommendations, and its deficiency and toxicity symptoms in farm animals have already been described in a previous opinion of the Scientific Committee on Animal Nutrition (SCAN) (EC, 2003a); the maximum levels authorised for total copper in feedingstuffs are derived from that opinion. To the knowledge of the FEEDAP Panel, there is no additional relevant information that may lead it to modify the SCAN opinion. Cupric chelate of amino acids hydrate is currently authorised as a nutritional additive under the functional group Compounds of trace elements to be used in feed for all animal species/categories. The applicant is asking the re-evaluation of that compound. The FEEDAP Panel reviewed the potential relation between the copper supply of animals and the development of antibiotic resistance in bacteria. The Panel also considered the Maximum Residue Limits (MRLs) set for copper in products of animal origin 11 resulting from the use of copper as pesticide, in the light of the use of copper in animal nutrition. The risk assessment is presented in a previous opinion of the FEEDAP Panel on copper sulphate pentahydrate (EFSA, 2012), and therefore not repeated in the current document. A compilation of risk assessments carried out on copper and its compounds, including opinions from EFSA panels other than the FEEDAP Panel, can be found in Appendix B. A list of authorisations of copper compounds in the EU, other than as feed additive, is reported in Appendix C. EFSA commissioned two studies, from which technical reports have been delivered; information from these reports has been used in this opinion. One of the studies was on selected trace and ultratrace elements in animal nutrition by the University of Gent (Belgium) (Van Paemel et al., 2010); copper was included in this study. The other study concerned the pre-assessment of the environmental impact of zinc and copper used in animal nutrition (Monteiro et al., 2010). 2. Identity and characterisation For compounds of trace elements, the element itself is considered the active substance. Cupric chelate of amino acids hydrate has the chemical formula Cu(x) 1 3 nh 2 O, where x = anion of any amino acid derived from hydrolysed soybean protein, molecular weight not exceeding 1500 Da Manufacturing process The manufacturing process comprises an acid hydrolysis of soybean protein under heat, followed by chelation with copper (from cupric oxide). The process is optimised to maximise the release of free amino acids. 12 The final additive is obtained by adding cellulose (44 45 %) and calcium carbonate (18 19 %) to the slurry and drying. 11 Commission Regulation (EC) No 149/2008 of 29 January 2008 amending Regulation (EC) No 396/2005 of the European Parliament and of the Council by establishing Annexes II, III and IV setting maximum residue levels for products covered by Annex I thereto. OJ L 58, , p Supplementary Information January

9 The applicant stated that the source soybean is of conventional, non-genetically modified, origin, either contractually sourced, or based on an approved geographic region, is used in the production of the additive Characterisation of the copper compound The copper content of five batches of the slurry 14 ranged from 8.95 to 9.17 % (w/v). 15 The applicant estimated the molecular weight of the additive to be Da. Analytical data on one sample showed that 98 % of the chelated material had a molecular weight <1500 Da and 2 % a molecular weight >1500 Da. 16 The lysinoalanine content of five batches of the hydrolysate was below 5 mg/100 g protein. 17 The content of heavy metals (lead, cadmium and mercury), fluorine and arsenic analysed in the slurry complied with EU legislation, as did the dioxins and the sum of dioxins plus dioxin-like PCBs (all values derived from three batches) Characterisation of the additive The final product is specified to contain a minimum of 10 % copper, as confirmed by analysis of five batches ( %). 19 Proximate analysis of the same five batches revealed a content of 3.5 % moisture, 21.1 % crude protein, 1.4 % lipids, 11.9 % crude fibre and 37 % ash. 20 The product also contains per kilogram additive: 56.5 g calcium, 4.7 g sodium, 3.3 g potassium, 1.25 g sulphur and 0.08 g phosphorous (one batch). Heavy metals (lead, cadmium and mercury), fluorine and arsenic complied with the limits set in EU legislation, as did dioxins and the sum of dioxins plus dioxin-like PCBs (all values derived from five batches). 21 The nickel content (five batches) was between 3.05 and mg/kg. 22 In five batches Escherichia coli O157.H7 and Salmonella (25 g sample of the product) were absent Physical state of the additive The product is a coarse granular powder with a slight characteristic odour. It is reported to be insoluble in water. Bulk density is kg/m 3. Sieve analysis of three batches identified that 1 2 % (w/w) of particles had a diameter below 180 µm. 24 At the request of the FEEDAP Panel sieve analysis was repeated in three other batches, and revealed the fraction below 75 µm to be % (w/w). 25 No data on dusting potential were provided Stability No data were provided for the cupric chelate amino acids hydrate, an organic copper compound. Information on the maintenance of the specific copper bonds in the chelates would be valuable. The FEEDAP Panel recognises the analytical difficulties in demonstrating stability of this specific bond and notes that the active substance is also unlikely to disappear in these products. 13 Supplementary Information/March Slurry means the intermediate product of copper chelation with the protein hydrolysate. 15 Supplementary Information/January Supplementary Information/January Supplementary Information/June Supplementary Information/January Technical Dossier/Section II/Annex II_2. 20 Technical Dossier/Section II/Annex II_2a and Annex II_2b. 21 Technical Dossier/Section II/Annex II. 22 Supplementary Information/March Technical Dossier/Section II. 24 Technical Dossier/Section II/Annex II and Supplementary Information/March Supplementary Information/March

10 Microbial decomposition could be excluded since Enterobacteriaceae, moulds and yeasts were present at <10 CFU/g and Salmonella was absent in 25 g after 10, 15 and 18 months storage Homogeneity The potential homogeneous distribution of the additive was predicted using a statistical approach (Jansen, 1992) in the case of a complete feed for chickens for fattening. The coefficient of variation was 11%. 27 However, the FEEDAP Panel notes that this method has been developed to test the working accuracy of mixing equipment and its applicability to test homogeneity has not been demonstrated Physico-chemical incompatibilities in feed Based on current knowledge, no incompatibilities resulting from the use of copper in compound feed are expected, other than those widely known and considered by feed manufacturers when formulating diets Conditions of use The copper compound under application, cupric chelate of amino acids hydrate, is intended to supply copper in final feed for all animal species/categories up to a maximum total content of 170 mg Cu/kg complete feedingstuffs for piglets (up to 12 weeks) and 25 mg Cu/kg for other pigs; 15 mg Cu/kg complete feedingstuffs for bovine before the start of rumination (milk replacers and other complete feedingstuffs) and 35 mg Cu/kg for other bovine; 15 mg Cu/kg complete feedingstuffs for ovine; 50 mg Cu/kg complete feedingstuffs for crustaceans; 25 mg Cu/kg complete feedingstuffs for fish; and 25 mg Cu/kg complete feedingstuffs for other species Evaluation of the analytical methods by the European Union Reference Laboratory (EURL) EFSA has verified the EURL report as it relates to the methods used for the control of the copper (seven compounds, including cupric chelate of amino acids hydrate) in animal feed. The Executive Summary of the EURL report can be found in the Appendix A. 3. Safety 3.1. Safety for the target species Tolerance of target animals Huge differences in the maximum tolerable copper concentrations between animal species exist (e.g. 500 mg/kg for rodents; 250 mg/kg for poultry, pigs and horses; 100 mg/kg for fish; 40 mg/kg for cattle, 15 mg/kg for sheep) (NRC, 2005). Therefore, considering the maximum copper content in feed set by Regulation (EC) No 1334/2003, it cannot be expected that all animal species would share the same margin of safety for a copper-containing additive. Consequently, the theoretical margin of safety (maximum tolerable concentration/maximum content authorised) of a safe copper-containing additive could be 10 in poultry and pigs (250/25), 4 in fish (100/25), 1.1 in cattle (40/35) and 1 in ovines (15/15). Since these margins of safety are not substance-specific, this consideration neglects the influence of potential differences in the bioavailability of copper from different sources. However, according to a literature review comparing the relative bioavailabilities of different sources of copper, including copper lysine and copper methionine, in poultry, pigs and ruminants, there was no evidence that the bioavailability of copper from organic forms is higher than the bioavailability from copper sulphate (Jongbloed et al. 2002). The conclusions of this review are supported by more recent publications cited in Section Consequently, the FEEDAP Panel does not expect the tolerance 26 Supplementary Information/March Technical Dossier/Section II. 10

11 of target animals to cupric chelate of amino acids hydrate to differ from tolerance to copper sulphate, when used up to the maximum authorised content in feed Microbial studies The antimicrobial resistance to copper from the additive was tested against five tetracycline-sensitive microbial strains as recommended by the FEEDAP Guidance on Microbial Studies (EFSA, 2008). The minimum inhibitory concentration for all five strains was above 50 mg Cu/L, the highest tested copper concentration. 28 The influence of copper in animal nutrition on the development of antibiotic resistance in bacteria has been reviewed and discussed in the scientific opinion on copper sulphate pentahydrate (EFSA, 2012) Conclusions on safety for the target species The FEEDAP Panel concludes that the source of copper under application is safe for all animal species/categories up to the maximum total copper content authorised in feed Safety for the consumer Metabolic and residue studies Copper is absorbed from the diet in the upper jejunum by active and passive processes, stored in the liver and kidney, secreted in the bile and excreted in faeces. Copper excretion via the kidneys is quantitatively insignificant if complex-forming substances are not administered (thiomolybdate from oral molybdenum in ruminants, dimethyl cysteine). Copper status is not easy to determine, and the homeostatic mechanisms that control copper distribution and metabolism are not completely understood. Copper interacts with other divalent cations, such as calcium, iron and zinc, for gastrointestinal absorption and metabolism. Absorption and availability may be influenced by the carbohydrate content of the diet, with reduced availability by phytate containing diets or those containing fructose (EC, 2003b). The SCAN delivered an opinion on copper (EC, 2003a) in which the metabolism and tissue deposition of copper were reviewed. Other reviews are available from McDowell (2003) and Suttle (2010). The distribution of total copper in the body varies with species, age and copper status of the animal. In general, levels in newborn and suckling animals are higher, then a steady decline during growth until adult values are reached. The main target organ for copper deposition is the liver. Other edible tissues containing high concentrations of copper are the heart, brain and kidney. Lower levels are found in muscle. Liver and kidney copper concentrations are related to dietary intake, whereas muscle is less affected. Copper is generally present in very low amounts in milk and this is not influenced by dietary supplementation levels. Clearance is higher in poultry than in mammals; the copper concentration in eggs is generally low. In fish, copper is primarily stored in the liver Organic vs. inorganic copper compounds The effects of trace mineral fortification of the diet in dairy cattle were studied by Nocek et al. (2006). This study included 573 cows, which were balanced by parity and 305-day mature equivalent at dryoff. Zinc, manganese, copper and cobalt in organic form, in the form of sulphates and as a combination of organic and inorganic forms with zinc and copper were supplied at dose levels up to 100 % of the NRC (2001) recommendations. Liver biopsies were collected from the same 30 cows per treatment at three times. Liver copper concentration increased with time (P< 0.01) and was higher in cows supplemented with organic copper and the combination of organic and inorganic copper sources than in those receiving the sulphate form, indicating that the liver responds to copper source. In a study with 300 multiparous dairy Holstein cows, treatments with different forms of dietary copper were given from 21 days prior to calving until 250 days of lactation (Ballantine et al., 2002). Diets 28 Technical Dossier/Section III/Annex III-1. 11

12 either contained only inorganic copper levels, or part of the copper was replaced by organic copper. Liver samples were taken by biopsy prior to initiation of treatments and at approximately 18 weeks post calving. No significant differences could be found. In another study in dairy cows (Siciliano-Jones et al., 2008), 250 multiparous and primiparous cows were assigned to groups receiving different forms of trace elements from approximately 70 days prepartum. Treatments consisted of supplemental zinc, manganese, copper and cobalt provided in sulphate form and 360 mg zinc, 200 mg manganese, 125 mg copper per animal and day in organic form. There was no effect of treatment on copper content of the liver. A study was conducted by supplementing the diet of laying hens with a combination of zinc, manganese and copper from organic and inorganic sources (Mabe et al., 2003). Addition of zinc, manganese and copper did not increase their concentrations in egg yolk irrespective of source Copper deposition in tissues and products of animal origin Following a request from EFSA, the applicant performed a literature search for peer-reviewed articles published between 2003 and November 2011 on potential differences in the deposition in edible tissues and products of copper from inorganic and organic sources. 29 The databases used for this research were Medline, Food Science and Technology, Agricola, Biological Abstracts, Embase, Medline preprints and Life science collections, and included the applicant s own database. The keywords applied were reported. Since the bioavailability and retention of copper can vary between different organic forms, only the studies in which copper chelate of amino acids hydrate was administered were considered to compare the copper edible tissue/animal products deposition with that of copper sulphate as standard inorganic copper source. In the case of liver copper deposition, data from three studies on cows, two studies on steers, one on pigs and four on poultry were submitted. In cattle for fattening, the supplementation of feed with copper (10 mg/kg DM) from 50 % sulphate and 50 % chelated amino acid hydrate did not result in different copper liver levels than those obtained after supplementation with the equivalent dose of copper sulphate only (Ahola et al., 2004). A study on dairy cows showed that oral doses of solutions containing the equivalent of 150 mg Cu/day, either from sulphate or from chelated amino acid hydrate, resulted in significantly different copper deposition in liver compared with control; however, there was no difference in copper liver concentration between cows treated with copper sulphate pentahydrate or copper amino chelate; the rate of copper deposition in liver was influenced by the initial copper concentration in liver (Balemi et al., 2010). In ovariectomised beef cows the supplementation of feed with copper (10 mg/kg DM) from chelated amino acid hydrate or copper sulphate did not lead to significantly different liver levels of copper (Ahola et al., 2005). No significant differences in liver levels of copper were found between cows given an inorganic source (copper sulphate or tribasic copper chloride) and those given copper from a chelated amino acid hydrate at either 5 mg Cu/kg (Mullis et al., 2003) or 10 mg Cu/kg (Arthington et al., 2003). No significant differences in the levels of copper in liver were reported in pigs for fattening given copper from copper sulphate or from chelated amino acid hydrate (0, 10, 30 and 50 mg/kg feed) (Hernandez et al., 2009). In laying hens fed diets supplemented 150 or 250 mg Cu/kg from either copper sulphate or chelated amino acid hydrate, the response in terms of liver copper levels was not significantly different (Idowu et al., 2006). Similarly, studies using a copper dose of 8 mg/kg feed (Ao et al., 2009; Aksu et al., 2010) from copper sulphate or from chelated amino acid hydrate did not result in a different pattern of copper liver deposition in chickens for fattening. Only in the study of Jegede et al. (2011), in which broilers diet was supplemented with copper at with copper supplementation levels of 50, 100 and 150 mg copper from copper sulphate or from chelated amino acid hydrate, did the highest element dose result in significantly higher copper liver deposition independent of source. However, this level of 29 Supplementary Information/March

13 copper feed supplementation (150 mg/kg) exceeds by six times the maximum EU authorised total copper level in complete feed. For kidney, data from only one study on pigs were submitted. No differences in kidney copper levels depending on the source of copper used were observed at the feed supplementation levels tested (0, 10, 30 and 50 mg Cu/kg feed) (Hernandez et al., 2009). For muscle copper deposition two studies in broiler chickens were cited. Petrovič et al. (2010) supplemented the diet with 0, 2.5 or 5 mg Cu/kg feed while Jegede et al. (2011) supplied chickens for fattening with supranutritional copper levels (50, 100 and 150 mg Cu/kg) from copper sulphate and chelated amino acid hydrate. Neither study found any differences in tissue deposition between the sources. Similarly, no difference in muscle copper concentrations in ewes was found when copper was given in the form of sulphate or chelated amino acid hydrate (Pal et al., 2010). The overall equivalent bioavailability of copper from both sources in terms of copper deposition is also supported by data from the tibia in chickens for fattening (Bao et al., 2007; Jegede et al., 2011). Regarding animal products, no difference in copper levels in eggs were observed between hens fed diets supplemented with copper sulphate or chelated amino acid hydrate at levels that did not exceed the maximum authorised total element level (Mabe et al., 2003) or with levels highly exceeding it (Idowu et al., 2006). Data from one study carried out with dairy cows (total dietary copper 13.8 mg/kg) did not show any difference in milk copper levels depending on copper source (Kinal et al., 2007). Based on the review of scientific literature dealing with use of cupric chelate of amino acid hydrate, the FEEDAP Panel does not consider the tissue/product deposition of copper from the additive under assessment to be any different from that of copper sulphate, which is a standard inorganic copper compound widely used in animal nutrition. Consequently, the use of the additive cupric chelate of amino acid hydrate in feed is not expected to modify the current consumer exposure to copper. The amino acids from the additive will be absorbed, enter the amino acid pool of protein and be metabolised accordingly Assessment of consumer safety A tolerable upper intake level (UL) for copper of 5 mg/day for adults and 1 mg/day for toddlers (one to three years) was defined by the SCF (EC, 2003b). This figure was derived from an overall no observed adverse effect level of 10 mg Cu/day identified in the study by Pratt et al. (1985) (daily single dose levels administered only to seven male adult volunteers for 12 weeks, and serum liver markers as endpoints), applying an uncertainty factor of 2 for potential variability in the normal population. This UL value has been consistently used in the assessments of copper in different forms by the following EFSA scientific panels: the NDA Panel (EFSA, 2006), the AFC Panel (EFSA, 2008b), the FEEDAP Panel (EFSA, 2008a, 2009a, 2012), the ANS Panel (EFSA, 2009b) and the CEF Panel (EFSA, 2010). Studies on copper dietary intakes in industrialised countries did provide comparable results. Mean dietary copper intakes by adults in different European countries have been estimated to be within a range of about mg/day (Van Dokkum, 1995; EC, 2003b; Sadhra et al., 2007; Rubio et al., 2009; Turconi et al., 2009). Based on 11 independent, peer-reviewed surveys considering only analytically confirmed copper (n = 849) in Belgium, Canada, the UK and the USA, the mean copper intakes for men and women were estimated to be 1.48 (95th percentile 2.87) and 0.92 (95th percentile 2.18) mg/day, respectively (Klevay, 2011). A recent analytical study of Catalonian diets showed a copper intake of 1.2 mg/day (Domingo et al., 2012). It has been suggested that calculated copper intakes are overestimated when only food composition tables are used in nutrition surveys (Klevay, 2012). 13

14 Among edible tissues of animal origin, the highest concentration of copper is found in liver (the main deposition organ), followed by kidney and muscle. Among products of animal origin, milk shows the lowest values (for quantitative figures, see EFSA, 2012). Since the supplementation of animal feed with copper-containing compounds has not essentially changed over the last decade, it is reasonable to assume that food of animal origin recorded in the above-mentioned consumption surveys originated from animals fed copper-supplemented diets and showing copper concentrations of tissues and products in the range mentioned above. The continued use of cupric chelate of amino acids hydrate in animal nutrition would not modify consumer exposure to copper Conclusions on the safety for the consumer Consumption surveys indicate that the mean dietary copper intake by adults in Europe is between 1.0 and 2.0 mg/day. These data include copper from foodstuffs of animal origin. Since the supplementation of animal feed with copper-containing compounds has not essentially changed over the last decade, no change in the contribution of foodstuffs originating from supplemented animals to the overall copper intake of consumers is expected. The use of the cupric chelate of amino acids hydrate in animal nutrition would not lead to different copper concentrations in food of animal origin compared with the standard authorised copper sulphate and would, therefore, not modify the exposure of consumers to dietary copper. The FEEDAP Panel concludes that the cupric chelate of amino acids hydrate additive is safe for consumers when used as a supplemental source of copper up to the maximum authorised levels for total copper in feedingstuffs Safety for the users/workers No specific studies have been provided by the applicant. Some copper compounds are recognised as skin and eye irritants. The additive should be considered as a skin and eye irritant and, owing to its amino acid/peptide component, as a skin/respiratory sensitiser Safety for the environment During the use of copper-containing feed additives, copper is unavoidably released into the environment. When used in livestock feed, copper excreted in the faeces will enter the soil environment when the faeces are applied as fertiliser to the land in the form of manure, slurry or litter. This may present two main potential risks: copper accumulation within the topsoil to concentrations posing potential toxic risk to soil organisms; leaching of copper from the soil to surface waters in concentrations posing potential toxic risk to organisms resident in the water column and bottom sediments. When used in aquaculture, trace elements such as copper may be released directly to the broader aquatic environment around an aquaculture facility or be taken up by fish and then excreted into the environment. As stated in the EFSA technical guidance for assessing the safety of feed additives for the environment (EFSA, 2008c), the compartment of concern for fish farmed in cages is assumed to be the sediment, whereas for fish farmed in land-based systems the effluent flowing to surface water is considered to pose the main environmental risk. EFSA commissioned a study on the environmental impact of zinc and copper used in animal nutrition (Monteiro et al., 2010). The results of this study were used as the basis for the present opinion. To assess the potential risks from copper used as additive in feed for terrestrial animals, a model that integrates the physicochemical and hydrological processes that determine the accumulation and leaching of metals in soil was used. Input rates of metals resulting from the use of feed additives and the spreading of animal manure on the land were based on the maximum allowable metal contents of 14

15 feed additives for different livestock types and the maximum allowable rates of nitrogen input of 170 kg/ha per annum. The assessment is based on the worst case assumption that the total amount of additive consumed will be excreted. Concentrations in surface water (as dissolved metal) and sediment (as total sediment metal) were calculated based upon the Forum for the Coordination of Pesticide Fate Models and their Use (FOCUS) scenario methodology and taking into consideration the speciation in the environment. More specific information on the parameterisation and assumptions made is given in the report. The predicted no-effect concentrations (PNECs) for the different compartments were calculated following the same methodologies as those presented in the EU risk assessment report for copper by correcting for bioavailability based on the assumed soil and water chemistry of the different scenarios. Likewise, it was decided to use the added PNEC approach for copper. The environmental risks of copper arising from aquaculture were assessed using the exposure models recommended in the technical guidance (EFSA, 2008d). The estimated concentrations in surface water resulting from the use of copper as feed additive for different fish species farmed in raceways, ponds, tanks and recirculation systems and the estimated concentration in sediment arising from the use of feed additives in sea cages were all below the PNEC and therefore do not give rise to concern. Concerning the terrestrial environment, potential risks to soil organisms have been identified as a result of application of piglet manure. However, of the nine scenarios in which a potential risk was identified, only two have local significance for pig production. Levels of copper in other types of manure are too low to create a potential risk within the timescale considered. For the water compartment, none of the scenarios resulted in the PNEC being exceeded when corrected for bioavailability. However, predicted concentrations of metals in the sediments of receiving waters, derived from the erosion of metal-enriched particles and transport in drainage and runoff, responded significantly to increases in metal inputs resulting from manure application. Potential risks were predicted within 50 years for several scenarios (i.e. R3, R1, D6, D5, D2 and D1). In two scenarios (D2 and R3) ), all manure types were predicted to result in exceedances. The predictions of the D2 scenario are of particular note as it is a cracking clay soil of the type vulnerable to bypass flow during events and thus potentially to extensive transport of particles to drainage as is simulated in that scenario. In the view of the FEEDAP Panel, these findings should be treated with caution as further model validation is needed and refinements are feasible, e.g. by taking into account the surface water chemistry of the locations of the FOCUS scenarios, more updated bioavailability models, resuspension and washout of deposited sediment and chemical transformation of trace elements in the sediment following deposition (i.e. formation of acid-volatile sulphide and metal sulphides). The extent to which copper-resistant bacteria contribute to the overall antibiotic resistance situation cannot be quantified at present (EFSA, 2012) Conclusions on safety for the environment Potential risks to soil organisms have been identified as a result of the application of piglet manure. Levels of copper in other types of manure are too low to create a potential risk within the timescale considered. There might also be a potential environmental concern related to the contamination of sediment owing to drainage and the run-off of copper to surface water. In order to draw a final conclusion, further model validation is needed and some further refinement to the assessment of copper-based feed additives in livestock needs to be considered, for which additional data would be required. The use of copper-containing additives in aquaculture, up to the maximum authorised copper level in feeds, is not expected to pose an appreciable risk to the environment. The extent to which copper-resistant bacteria contribute to the overall antibiotic resistance situation cannot be quantified at present. 15

16 4. Efficacy Cupric chelate of amino acids for all species The use of cupric chelate of amino acids hydrate in animal nutrition is well documented in scientific literature. It is recognised as an efficacious source of copper to meet animal requirements (Baker and Ammerman, 1995). The efficacy of the cupric chelate of amino acids hydrate was further demonstrated in two studies provided by the applicant one in dairy cows (Ballantine et al., 2002) and one in laying hens (Mabe et al., 2003). 5. Post-market monitoring The FEEDAP Panel considers that there is no need for specific requirements for a post-market monitoring plan other than those established in the Feed Hygiene Regulation 30 and Good Manufacturing Practice. CONCLUSIONS AND RECOMMENDATIONS CONCLUSIONS Cupric chelate of amino acids hydrate is safe for all animal species/categories up to the authorised maximum of total copper content in complete feed. Consumption surveys include copper from foodstuffs of animal origin. Since the supplementation of animal feed with copper-containing compounds has not essentially changed over the last decade, no change in the contribution of foodstuffs originating from supplemented animals to the overall copper intake of consumers is expected. No concerns for consumer safety are expected from the use of cupric chelate of amino acids hydrate in animal nutrition, which would substitute for other copper sources. The additive should be considered as a skin and eye irritant and, owing to its amino acid/peptide component, as a skin/respiratory sensitiser. Potential risks to soil organisms have been identified as a result of the application of piglet manure. Levels of copper in other types of manure are too low to create a potential risk within the timescale considered. There might also be a potential environmental concern related to the contamination of sediment owing to drainage and the run-off of copper to surface water. In order to draw a final conclusion, further model validation is needed and some further refinement to the assessment of copper-based feed additives in livestock needs to be considered, for which additional data would be required. The use of copper-containing additives in aquaculture up to the authorised maximum of total copper content in complete feeds is not expected to pose an appreciable risk to the environment. The extent to which copper-resistant bacteria contribute to the overall antibiotic resistance situation cannot be quantified at present. Cupric chelate of amino acids hydrate is recognised as an efficacious source of copper to meet animal requirements. RECOMMENDATIONS The description of the product proposed by the applicant should be amended as follows: Cu(x) 1 3 nh 2 O (x= anion of any amino acids derived from acid hydrolysed soya protein). At least 90 % of the molecules should have a molecular weight not exceeding 1500 Dalton. Since the copper compounds are not available for control, and valuable information to the user of the product can only be given for the final product, the FEEDAP Panel proposes to evaluate the possibility of introducing the formulated product in the register of feed additives. 30 Regulation (EC) No 183/2005 of the European Parliament and of the Council of 12 January 2005 laying down requirements for feed hygiene. OJ L 35, , p

17 GENERAL REMARK More recent findings on the copper requirements of animals indicate the potential to considerably reduce the current maximum content for dietary copper without affecting animal health and welfare and the productivity of animal husbandry. A reduction in the maximum content of copper would decrease the copper load in the environment. The FEEDAP Panel stresses the need for analytical methods to detect the organic compounds of trace elements in feed, independent from the trace element background. DOCUMENTATION PROVIDED TO EFSA 1. Dossier Copper Amino Acid Chelate, Hydrate (Availa Cu) for all animal species. July Submitted by Zinpro Animal Nutrition Inc. 2. Dossier Copper Amino Acid Chelate, Hydrate (Availa Cu) for all animal species.. Supplementary information. March Submitted by Zinpro Animal Nutrition Inc. 3. Dossier Copper Amino Acid Chelate, Hydrate (Availa Cu) for all animal species.. Supplementary information. June Submitted by Zinpro Animal Nutrition Inc. 4. Dossier Copper Amino Acid Chelate, Hydrate (Availa Cu) for all animal species.. Supplementary information. January Submitted by Zinpro Animal Nutrition Inc. 5. Evaluation report of the European Union Reference Laboratory for Feed Additives on the methods(s) of analysis for Copper (E4). 6. Comments from Member States received through the ScienceNet. REFERENCES Ahola J K, Baker DS, Burns PD, Mortimer RG, Enns RM, Whittier JC, Geary TW and Engle TE, Effect of copper, zinc, and manganese supplementation and source on reproduction, mineral status, and performance in grazing beef cattle over a two-year period. Journal of Animal Science, 82, Ahola JK, Engle TE and Burns P D, Effect of copper status, supplementation, and source on pituitary responsiveness to exogenous gonadotropin-releasing hormone in ovariectomized beef cows. Journal of Animal Science, 83, Aksu DS, Aksu T, Özsoy B and Baytok E, The effects of replacing inorganic with a lower level of organically complexed minerals (Cu, Zn, and Mn) in broiler diets on lipid peroxidation and antioxidant defense systems. Asian-Australian Journal of Animal Science, 23, Ao T, Pierce JL, Power R, Pescatore AJ, Cantor AH, Dawson KA and Ford MJ, Effects of feeding different forms of zinc and copper on the performance and tissue mineral content of chicks. Poultry Science, 88, Arthington JD, Pate FM and Spears JW, Effect of copper source and level on 23 performance and copper status of cattle consuming molasses-based supplements. Journal of Animal Science, 81, Baker DH and Ammerman CB, Copper bioavailability. In: Bioavailability of nutrients: amino acids, minerals, and vitamins. Ammerman CB, Baker DH and Lewis AJ. Academic Press, San Diego, CA, Balemi SC, Grace ND, West DM, Smith SL and Knowles SO, Accumulation and depletion of liver copper stores in dairy cows challenged with a Cu-deficient diet and oral and injectable forms of Cu supplementation. New Zealand Veterinary Journal, 58,