Copper and zinc in diets of growing pigs

Copper and zinc in diets of growing pigs New insights in requirements Paul Bikker and Age Jongbloed

Outline Introduction Copper and zinc as essential trace element Present: recommendations, allowance and practice Two studies into zinc and copper requirements Objectives Experimental design Results and discussion Conclusions Practical implications

Copper and Zinc as essential trace elements Numerous physiological functions of zinc and copper Component of >200 metalloenzymes Zn, required for DNA transcription (zinc fingers) almost all metabolic processes depend on zinc Cu, red blood cells, iron metabolism, anti-oxidant Consequences of Cu and Zn-deprivation Loss of appetite reduced growth rate (Zn) Skin abnormalities (parakeratosis, thickening, lesions) (Zn) Bone development, growth, reproduction (Cu, Zn) Cu-deprivation only after experimental induction

Cu and Zn: recommendations in mg/kg Source Zinc Copper Weaned pigs BSAS, 2003 +100 +6.0 GfE, 2008 80-100 6.0 NRC, 1998 100 80 6.0 5.0 INRA, 1989 100 10 GF pigs BSAS, 2003 +100 60 +6.0 GfE, 2008 50-60 4.0-5.0 NRC, 1998 60 50 4.0 3.0 INRA, 1989 100 10

Zinc and copper allowance, max. levels Zinc Copper EC 1987 2003 PDV 2000 2003 EC 2004 EC 1987 2003 EC total content, PDV added via premix From ingredients: 30-40 mg Zn, 5-8 mg Cu per kg PDV 2000 2003 EC 2004 Pigs < 12 wk 250 +100 150 175 +160 170 Pigs 12-16 wk 250 + 70 150 175 +130 25 Pigs >16 wk 250 + 60 150 35 + 15 25 Breeding pigs 250 + 65 150 35 + 20 25

Zinc in Complete feed for Pigs C onte nt (m g/k g) 300 250 200 150 100 50 250 200 150 100 50 Num ber of sam ples 0 2001 2002 2003 2004 2005 2006 2007 2008 2009 0 limit Avg Median 90per Number trend avg y= 3.6x + 96 R 2 = 0.72 N = 1228 Adamse et al., 2011

Intermediate conclusions 200 Copper in Compleet feed for Piglets (< 12 wk) 45 Content (mg/kg) 180 160 140 120 100 80 60 40 20 40 35 30 25 20 15 10 5 Number of samples 0 2001 2002 2003 2004 2005 2006 2007 2008 2009 0 limit Avg Median 90per Number trend avg y= 0.5x + 141 R 2 = 0.01 N = 211 Adamse et al., 2011

Adamse et al., 2011

Copper and zinc balance BW, kg Cu, mg/kg Zn, mg/k g Cu, g Zn, g Nursery 8-25 165 130 4.2 3.3 GF-pigs 25-120 25 130 6.2 32.1 Retention 8-120 1.2 22 0.13 2.2 Excretion 10.2 33.2 Excretion, % 98.7 93.7 Not included Grower diet 2 wk +140 +2.4 Grower diet 4 wk +140 +5.9

Intermediate conclusions Recommendations poorly documented and applied Dietary copper and zinc levels close to EC maximum >90% excretion in manure Aim: reduction in excretion without loss of animal performance and health Required: reliable recommendations for practical use Assumption: highest requirements in weaned piglets

Experimental objectives Effect of dietary zinc and copper levels on performance and health in young growing pigs Zinc and copper requirements Nutrient digestibility Effect of microbial phytase Effect of organic acid inclusion (only zinc) Effect of growth promoting level of copper

Experiment zinc, treatments Treatment Added Zn mg/kg Total Zn Analysed Zn Phytase FTU/kg Lactic acid, % 1 0 30 33-0.8 2 15 45 47-0.8 3 30 60 61-0.8 4 45 75 74-0.8 5 60 90 86-0.8 6 100 130 127-0.8 7 15 45 46 500 0.8 8 15 45 47 500 - Cu, added 160 mg/kg Phytase, analysed, 300 (intrinsic) and 1100 FTU/kg

Experiment zinc, design 720 weaned piglets, females and castrates 8 Treatments x 10 pens x 9 piglets 2 wk adaptation / depletion, low zinc diet (+10 ppm) 1 piglet sacrificed or excluded from each pen 6 wk main period: 14-56 d post weaning Water and feed ad libitum Practical diets, adequate nutrient levels Digestibility in 6 replicates

Experiment zinc, observations D14, 28, 42, 56: body weight, feed intake D34, 55: faecal samples digestibility D0, 14, 35, 56: blood samples ALP, Zn, Cu D14, 56: body tissues liver, bone tissue Daily: health, medical treatments, losses 3x / wk: faecal consistency D56: score of skin and legs

Results zinc, faecal consistency Piglets with soft faeces: wk 1 20% wk 6 <1% No effects of dietary treatments Zn, Phy, LA Note dietary treatments in wk 3-8 post weaning No obvious effects of low zinc in prestarter diet (10 ppm)

Results zinc, health T 1 T 2 T 3 T 4 T 5 T 6 P- value 1 Added Zn, mg/kg 0 15 30 45 60 100 Piglets, n 80 80 80 80 80 80 Skin lesions - Light, n 28 b 0 a 2 a 1 a 0 a 1 a <0.001 - Severe, n 16 b 0 a 0 a 0 a 0 a 0 a <0.001 Lesions of legs - Light, n 25 b 0 a 0 a 0 a 0 a 0 a <0.001 - Severe, n 24 b 0 a 0 a 0 a 0 a 0 a <0.001

Results zinc, feed intake

Results, body gain

Results zinc, body weight (BW) and homogeneity Added Zn (mg/kg) T 1 T 2 T 3 T 4 T 5 T 6 P-value 2 0 15 30 45 60 100 Model BW d 56, kg 32.7 a 39.5 b 39.8 b 39.7 b 39.9 b 39.5 b <0.001 SD BW d 56, kg 6.13 b 3.73 a 3.61 a 3.23 a 4.10 a 3.49 a <0.001

Results, feed conversion ratio

Results, serum alkaline phosphatase

Results, serum zinc concentration

Results, liver and bone zinc content

Results of phytase on digestibility (%) T 2 T 7 T 8 SEM 1 P- value 2 Phytase No Yes Yes Model Organic acid Yes Yes No Mean Ca 47.8 a 59.1 b 57.2 b 0.77 *** P 52.8 a 64.2 b 63.5 b 0.77 *** Cu -9.2 a -3.7 b -2.9 b 1.48 * Zn 0.9 a 10.1 b 15.2 c 1.18 ***

Estimates of zinc requirements Characteristic Linear-plateau max. breakpoint Performance ADG, g/d 676 ± 8.0 47 ± 1.5 FI, g/d 1119 ± 0.06 46 ± 1.5 FCR 1.65 ± 0.015 48 ± 2.5 Blood serum ALP D35, U/L 202 ± 5.1 63 ± 2.6 Zinc D35, µmol/l 12.6 ± 0.23 70 ± 2.6 ALP D56, U/l 169 ± 4.0 62 ± 2.1 Zinc D56, µmol/l 12.3 ± 0.31 67 ± 2.7 Liver, mg/kg Zinc in DM 167 ± 6.9 83 ± 8.5

Conclusions zinc Zinc requirements No added zinc: reduced intake and severe clinical signs of deficiency 46-48 mg/kg adequate for body gain and health 62-70 mg/kg adequate for blood parameters Substantial contribution of phytase to Zn supply No added value of organic acid inclusion

Experiment copper, treatments Treatment Added Cu mg/kg Total Cu mg/kg Analysed Cu mg/kg Phytase FTU/kg 1 0 7 7-8 - 2 3 10 9-11 - 3 6 13 12-13 - 4 9 16 14-15 - 5 12 19 18-20 - 6 18 25 24-7 160 165 165-170 - 8 0 7 7-8 500 Zn, added 45 mg/kg, analysed 70-75 Phytase, analysed, 165 (intrinsic) and 760 FTU/kg

Experiment copper, design 640 weaned piglets, females and castrates 8 Treatments x 10 pens x 8 piglets No depletion period 2 wk prestarter, 6 wk starter diets 8 wk experimental period: 1-56 d post weaning Water and feed ad libitum Practical diets, adequate nutrient levels Digestibility in 6 replicates

Experiment copper, observations D0, 14, 28, 42, 56: body weight, feed intake D27, 55: faecal samples digestibility D0, 28, 56: blood samples Zn, Cu, Hb, Ht D0, 56: body tissues liver, bile Daily: health, medical treatments, losses 3x / wk: faecal consistency D56: score of skin and legs

Results, feed intake

Results, body gain

Results copper, feed conversion ratio

Results copper in liver and bile

Effect of phytase on nutrient digestibility (%) T 1 - T 8 +500 FTU SEM P-value DM 83.4 84.3 0.20 0.003 Ca 42.0 47.6 1.81 0.044 P 47.4 51.7 1.64 0.084 Cu 22.1 6.7 2.38 <0.001 Zn -0.6 10.1 1.05 <0.001

Effect of high copper on growth performance T 1-6 7-25 mg T 7 165 mg SEM P-value BW D56, kg 38.4 41.1 0.55 <0.001 SD BW D56, kg 3.7 3.9 0.35 0.612 ADG, g/d 540 587 9.7 <0.001 FI, kg/d 0.88 0.94 0.015 <0.001 FCR 1.63 1.60 0.013 0.032

Effect of high copper, T 7 as % of T1-6

Effect of phytase on tissue levels T 1 - mg T8 +500 FTU SEM P-value Day 28 Copper, µmol/l 24.5 23.6 0.98 0.53 Zinc, µmol/l 10.0 12.8 0.44 <0.001 Day 56 Copper, µmol/l 26.0 27.7 1.35 0.39 Zinc, µmol/l 12.4 17.6 0.51 <0.001 Liver, mg/kg DM 20.5 25.6 2.84 0.22 Bile, mg/l 1.23 2.23 0.41 0.10

Effect of high copper on tissue levels T 1-6 7-25 mg T 7 165 mg SEM P-value Day 28 Copper, µmol/l 24.5 26.4 0.98 0.071 Zinc, µmol/l 10.0 11.0 0.44 0.028 Day 56 Copper, µmol/l 28.6 30.9 1.35 0.19 Zinc, µmol/l 12.8 12.6 0.51 0.72 Liver, mg/kg DM 28.8 105.2 2.84 <0.001 Bile, mg/l 2.6 7.4 0.41 <0.001

Effect of high copper (T7) on faecal consistency

Effects (ns) of withdrawal of high copper, T 7 (160/15) as % of T 5 (12/15 mg/kg)

Conclusions copper Copper requirements No effect of added copper on growth performance 7 mg/kg adequate for body gain and health 7-10 mg/kg adequate for blood parameters No consistent effect of phytase on copper supply Substantial effect of 160 mg Cu on growth performance and faecal consistency Numerical reduction in growth performance after withdrawal of high copper level

Copper and zinc balance, implications BW, kg Cu, mg/kg Zn, mg/kg Cu, g Zn, g Nursery 8-25 165 80 4.2 2.0 GF-pigs 25-120 25 80 6.2 19.8 Retention 8-120 1.2 22 0.13 2.2 Excretion 10.2 19.6 Excretion, % 98.7 89.7 Reduction, % 41% Not included Grower diet 2 wk +140 +2.4 Grower diet 4 wk +140 +5.9

Implications Reduction of zinc to 80 mg/kg 244 tons lower Zn excretion 30% reduction of annual accumulation Further reduction possible taking into account the phytase contribution More insight required in growth promoting effect of Cu; level, duration, withdrawal Validation of low copper content in GF pig diets

Copper in diet and liver (Jondreville et al., 2002)

Copper and zinc balance, future challenge? BW, kg Cu, mg/kg Zn, mg/kg Cu, g Zn, g Nursery 8-25 165 55 4.2 1.4 GF-pigs 25-120 15 50 3.7 12.4 Retention 8-120 1.2 22 0.13 2.2 Excretion 7.8 11.5 Excretion, % 98.3 83.7 Reduction, % 24% 65% Not included Grower diet 2 wk +140 +2.4 Grower diet 4 wk +140 +5.9

Acknowledgements We highly appreciate The financial contribution of the Product Board Animal Feed (PDV) and the Ministry of Economic Affairs, Agriculture and Innovation (EL&I). The input of their representatives E.R. Deckers (EL&I), H. Boelrijk, M.C. Blok, A. Cazemier, M. Hessing, C.A. Makkink, J. Michels, and A. van Wesel (PDV) Co-operation with staff of VIC Sterksel

Thank you for your kind attention