Essential Trace Minerals

for Exceptional Performance

Zinpro Performance Minerals® deliver proven benefits for Shrimp Performance, Health and Meat Quality

Trace minerals play numerous essential roles within cells and metabolic processes, which makes them vital for optimal nutrition and health of animals – including shrimp.

Research shows that supplementing shrimp diets with Zinpro Performance Minerals is essential for optimizing growth and improving animal wellness through modulation of the immune system. In addition, meat quality is improved, as measured by color and drip loss.

The source and availability of trace minerals is key to satisfy the needs of shrimp in an efficient and sustainable way throughout their life cycle.

Trace Mineral Benefits in Shrimp

Meat Quality Nervous System Zn, Mn, Cu, Cu Fe, Se

Muscle Disease Resistance Development Zn, Mn, Cu, Fe, Se Zn, Se, Cr, Cu

Exoskeleton Development Epithelial Tissues Zn, Mn, Cu Zn, Mn, Cu Larvae and Post-larvae Development Zn, Mn, Cu, Se Study 1

Comparison of Whiteleg Shrimp Response to Inorganic Minerals and Zinpro Performance Minerals® (0.5x)

Key Findings

• Zinpro Performance Minerals (ZPM) • Cumulative mortality of shrimp challenged supplemented at half the level of inorganic with Vibrio harveyi was significantly (P < 0.05) mineral sources resulted in numerically higher reduced when inorganic mineral sources were final body weight (Fig. 1). partially or completed replaced with ZPM (Fig. 2).

• Return on investment (ROI), as measured by • Furthermore, drip loss of peeled white shrimp additional income over feed cost, was 16% more was significantly (P < 0.05) lower for shrimp fed for shrimp fed ZPM at 0.5x rate as compared diets supplemented with ZPM (Fig. 3). to shrimp fed inorganic minerals. ROI for the combination of inorganic and ZPM was 11% more than inorganic alone.

Growth Performance Fig. 1

Shrimp Weight, gram (8 Weeks)

11.5

10.5 Inorganic Inorganic/ZPM ZPM, 0.5x

Source: Jintasataporn, O., Ward, T., Chumkam, S, Jintasataporn, O. 2015. The Efficacy of Mineral-Amino Acid Complex (Zn, Mn, Cu, Fe and Se) in Diets to Growth Performance, Immune Status and Meat Quality of White Shrimp, Litopenaeus vannamei. Aquac.Indones. 16:33-37 Study Criteria Health Fig. 2 Cumulative Mortality. P5 This study compared the efficacy of Zinpro Performance Minerals Cumulative Mortality, % (metal-amino acid complexes) with Inorganic Inorganic/ZPM ZPM, 0.5x inorganic minerals (sulfates) in the diet for whiteleg shrimp. Growth performance, resistance to bacterial 100 z z challenge Vibrio harveyi, and meat 80 y Inorganic quality were evaluated. y 60 y y Inorganic ZPM (sulfate) Inorg + ZPM (0.5x) Inorganic/Metal-AA 40 Zinc (ppm) 120 70 50 60 Manganese 20 60 40 20 30 Me tal-AA 0 .5X (ppm) Copper 32 22 10 16 0 (ppm) Iron 1 22 33 44 55 66 77 (ppm) 100 50 50 50 Days after challenge with Vibrio Harveyi Selenium Days after challenge with Vibrio harveyi (ppm) 0.3 0 0.3 0.15

Initial body weight: 4.4g Stocking density: 70 shrimp/m2 Meat Quality Fig. 3 Feeding: 4x per day Replications: 6 Feeding period: 8 weeks Peel Shrimp 4 Day Post-Refrigeration Salinity: 12ppt 4

35.5% CP/7.5% Fat 3 2.92z 25% fish meal, 3% shrimp meal, 2% squid meal, 20% soybean meal, 2 2.25y 2.03y 5% poultry by-product meal Drip Loss, % Drip Loss, 1 Location: 0 Kasetsart University, Thailand Inorganic Inorganic/ ZPM, 0.5x ZPM Study 2

Availa®Zn and Availa®Se Supplementation Improves Growth, Health and Meat Quality

Key Findings

• Partial replacement of inorganic zinc with and after boiling (data not shown). Availa Se Availa Zn significantly (P < 0.05) improved shrimp also significantly (P < 0.05) improved redness of weight (Fig. 1). shrimp meat frozen for 14 days (Fig. 3).

• Partial replacement of inorganic zinc and • Whole shrimp drip-loss and meat rancidity selenium with Availa Zn and Availa Se were both significantly (P < 0.05) reduced by significantly (P < 0.05) increased hemocyte count partially replacing inorganic Se with Availa Se and phenoloxidase activity, key biomarkers of (Fig. 3). Moreover, the effect of partially replacing shrimp immune response (Fig. 2). inorganic Zn with Availa Zn on drip-loss tended to be more evident when an inorganic source of • Modulation of the immune response by ZPM Se was used. resulted in reduced cumulative mortality of shrimp challenged by Vibrio harveyi. • Zn and Se accumulation significantly (P < 0.05) increased in the exoskeleton and hepatopancreas • Availa Se significantly (P < 0.05) improved for shrimp fed Availa Zn and Availa Se (data not redness of 72-hour chilled shrimp meat, before shown).

Growth Performance Fig. 1

Shrimp Weight, gram

Inorganic Zinc Availa Zn 12 11.6z 11.0y 10 9.6z 8.9y 8 7.5 7.3 6 5.8z 4.4 4.5 5.5y

4 0 2 4 6 8 Weeks

Source: Mihai Sun, Terry Ward, Orapint Jintasataporn, Claudia Silva 2018. Efficacy of Availa Zn and Availa Se for White Shrimp (Litopenaeus vannamei). International Symposium on the Feeding and Nutrition of Fish, June 3-7, 2018, Las Palmas, Gran Canaria, Spain Study Criteria Health Fig. 2 This study evaluated the effects of Phenoloxidase Activity partially replacing inorganic Zn and Units/min/mg protein Se with Zn from Availa Zn and Se 120 120 from Availa Se on whiteleg shrimp 100 z 100 z growth performance, health and 80 y 80 y meat quality. 60 60 40 40 Inorganic Inorganic Zinc Inorganic Inorganic Se + Inorganic Zn Availa Zna Inorganic Zn Availa Zn b Zinc + Availa Zn Se Availa Se Inorganic Se Inorganic Se Availa Se Availa Se Zinc sulfate (ppm) 120 70 120 70 Availa-Zn (ppm) 50 50 Meat Quality Fig. 3 Sodium selenite 0.30 0.30 0.15 0.15 (ppm) Availa-Se Shrimp Meat Color (ppm) 0.15 0.15 Boiled after 72-hours chilled a Availa-Zn zinc amino acid complex 24 b Availa-Se zinc-L-selenomethionine

a 18 z Initial body weight: 4g 2 12 Stocking density: 60 shrimp/m Feeding: 4x per day Redness (a*) 6 y Replications: 6

0 Feeding period: 8 weeks Inorganic Se Inorganic Se + Availa Se Salinity: 30ppt a Higher redness value (a*) indicates redder color (+60 for red to -60 for green) 36.5% CP/8% fat Drip Loss, % 25% fish meal, 3% shrimp meal Whole shrimp, after 72-hours chilled 2% squid meal, 20% soybean meal 7 z Inorganic Se 5% poultry by-product

5 y Inorganic Se + Availa Se Location: 3 Kasetsart University, Thailand

1 Inorganic Zinc Inorganic Zinc + Availa-Zn Study 3

Supplementation with Availa®Fe and Availa®Se are Key in Coloration of Whiteleg Shrimp

Key Findings

• Diets supplemented with Availa Se showed • Diet supplemented with 75ppm astaxanthin, improvements in survival rate (Fig. 1) and FCR 200ppm Availa Fe and 0.3ppm Availa Se (AsFeSe) (data not shown). significantly (P < 0.05) enhanced shrimp meat redness as indicated by the a* value and total • The diet containing astaxanthin (As), Availa Se carotenoid content of cooked shrimp shell and and Availa Fe significantly (P < 0.05) increased flesh (Fig. 3). SalmoFan™ was able to score an the hemocyte count and phenoloxidase activity increase in shrimp color from 23 to 29 in only (Fig. 2). four weeks with AsFeSe (data not shown).

Growth Performance Fig. 1

Survival Rate, %

100 z 90 y 80 y y y 70

Control As AsFe AsSe AsFeSe Health Fig. 2

Hemocyte Count Phenoloxidase Activity x106 cell/ml Unit/min/mg protein 8 600 z 500 z 6 400 yz y 4 xy 300 xy 200 x x 2 w wx 100 0 0 6 weeks 6 weeks

Source: Jintasataporn, O., Ward, T., Chalemlap, S., Chumkam, S. Iron amino acid complex (Availa Fe) and zinc-L-selenomethionine (Availa Se) in diets enhance cooked Pacific white shrimp (Litopenaeus vannamei) color. International Symposium on the Feeding and Nutrition of Fish, June 3-7, 2018, Las Palmas, Gran Canaria, Spain. Study Criteria Meat Quality Fig. 3 This study was conducted to investigate shrimp coloration and Control As AsFe AsSe AsFeSe immune response when astaxanthin was added to the diet alone or in The a* Value of Cooked Shrimp Fed with combination with either iron or 5 Different Experimental Diets for 6 Weeks selenium, or with both iron and selenium. 60 50 Astaxanthin Availa Fe Availa Se 40 z z Treatments z (ppm) (ppm) (ppm) y y y y 30 y y Control - - - xy xy x

a* Value x 20 x w As 75 - - 10 AsFe 75 200 - AsSe 75 - 0.3 0 2 weeks 4 weeks 6 weeks AsFeSe 75 200 0.3

2.5 Initial body weight: c.a. 6.5g 2.0 z z Stocking density: 10 shrimp/ z y yz 100L aquarium 1.5 z z x yz Feeding: 3x per day xy y 1.0 y w Replications: 4 x x 0.5 Feeding period: 6 weeks

Shell Carotenoid Content, (mg/g) Shell Carotenoid Content, 0 2 weeks 4 weeks 6 weeks 12.5% fish meal, 3% squid meal, 25% soybean meal, 7.5% soy protein 1.2 concentrate, 3% yeast, 10% wheat gluten, 22% wheat flour 1.0 z z 0.8 0.6 Location: y 0.4 y Kasetsart University, Thailand x x 0.2 z x x x x y y y y 0 Flesh Carotenoid Content, (mg/g) Flesh Carotenoid Content, 2 weeks 4 weeks 6 weeks Study 4

Zinpro Performance Minerals® Spare Carotenoids For Pigmentation Purposes

Key Findings

• Supplementation with ZPM can effectively Availa®Zn, 40ppm Mn as Availa®Mn and 20ppm and economically support the reduction of Cu as Availa®Cu to AxCx diet (25ppm astaxanthin astaxanthin while maintaining cooked shrimp 25ppm canthaxanthin) increased SalmoFan color, performance and health parameters (data score of shrimp shell back to that of control diet not shown). (75ppm astaxanthin).

• The reduction of astaxanthin from 75 to 25ppm • ZPM supplementation reduced lipid oxidation, in combination with 25ppm canthaxanthin had measured as milligram malonaldehyde per gram no effect on lipid oxidation (Fig. 1) but it reduced shrimp (Fig. 1) highlighting antioxidant functions meat and shell redness as indicated by a* value of TM as co-factors of key antioxidant enzymes. (Fig. 2) and SalmoFan™ scores (Fig. 3). • Results indicate that supplementation with ZPM • SalmoFan scores of shrimp shell dropped from (AxCx FSe TMx2 treatment) can boost shrimp 27 to 24.4 in only 3 weeks when fed 25ppm antioxidant capacity in only 3 weeks, allowing astaxanthin in combination with 25ppm astaxanthin to be diverted for pigmentation canthaxanthin and no ZPM. purposes instead of antioxidant functions (sparing effect) saving $66 US per ton of feed. • Supplementation with 100ppm Fe as Availa®Fe, 0.3ppm Se as Availa®Se, 100ppm Zn as

Meat Quality Fig. 1

Lipid Oxidation, Week 3

Ax AxCx AxCx FSe AxCx FSe TMx1 AxCx FSe TMx2

z 12 z

y y 8 y

4 mg Malonaldehyde/g Shrimp 0

Source: Jintasataporn et al., 2020. Trace Mineral Amino Acid Complexes (Availa Zn, Mn, Cu and Fe) and Zn-L-Selenomethionine (Availa-Se) in white leg shrimp (Litopenaeus vannamei) Diets Enhance Cooked Shrimp Color. Aquaculture America, February 9-12th, 2020, Hawaii. Study Criteria

Meat Quality Fig. 2 Trace minerals participate as cofactors of important enzymes Colorimeter a* Value of Shrimp, Week 3 related to the antioxidant defense mechanism. This study evaluated Meat, a* Shell, a* 22 their effect on sparing carotenoids for pigmentation purposes. 20

a Caratenoids , b 16 ppm ZPM , ppm

Treatment Availa Availa Availa Availa Availa 14 ASTX CANX Fe Se Zn Mn Cu Ax 75 ------12 AxCx 25 25 - - - - - 10 AxCx FSe 25 25 100 0.3 - - - Ax AxCx AxCx FSe AxCx FSe AxCx FSe TM1x TM2x AxCx FSe TMx1 25 25 100 0.3 50 20 10 AxCx FSe TMx2 25 25 100 0.3 100 40 20

a Wisdom pink 10%, Wisdom red 10%; ASTX = astaxanthin, CANX = canthaxanthin b ZPM were supplemented on top of basal diet that included inorganic premix (mg/kg diet) : 0.2 Co, 25 Cu, 1 I, 30 Fe, 30 Mn, 0.35 Se, 100 Zn. Meat Quality Fig. 3 Initial body weight: 7–8g SalmoFan™ Score, Week 3 Stocking density: 20 shrimp/300L tank Meat Shell Feeding: 3x per day 28 Replications: 5 Feeding period: 3 weeks 27 x x Salinity: 10-15ppt 26 b b z 25 b b z 38% CP/6% Fat 24 y 12.5% fish meal, 3% squid meal, 23 a 25% soybean meal, 7.5% fermented 22 soy meal, 3% yeast, 10% wheat gluten, 21 22% wheat flour Ax AxCx AxCx FSe AxCx FSe AxCx FSe TM1x TM2x Location: Kasetsart University, Thailand Study 5

A. Effect of Copper Source on Growth and Intestinal Microbial Communities of Whiteleg Shrimp

Key Findings

• Cu supplementation positively affected growth • Combination of Availa Cu + CuSO4 Cu (total of shrimp. 30ppm Cu) resulted in numerically better growth

® than Availa Cu at 0.5x rate, indicating higher Cu • Availa Cu at 0.5x rate (15ppm Cu) of CuSO4 supplementation may be required for maximized (30ppm Cu) maintained shrimp FBW (Fig. 1), shrimp performance. showing Availa Cu is more effective than inorganic Cu. • The reduction on Proteobacteria phylum which includes the Vibriocaceae family is related to • Marked trends in microbial phyla suggest overall shrimp health when a fed combination of microbial communities were being influenced CuSO and Availa Cu or Availa Cu at 0.5x rate. by copper source (Fig. 2a). 4

Growth Performance Fig. 1

Final Body Weight, gram

13.5

13.0 z

12.5 y yz 12.0

11.5 x

11.0

Control CuSO4 CuSO4 /AvCu AvCu 0.5x Health Fig. 2

2a. Percentage Distribution of Bacterial Phyla In L. vannamei Intestine

Proteobacteria Bacteroidetes Tenericutes Fusobacteria CKC4 Other Bacteria

100%

80%

60%

40%

20%

0% Control CuSO4 CuSO4 /AvCu AvCu 0.5x

Note: Vibrios are part of the Proteobacteria Phyla

Source: Yuan et al., 2019a. Effects of different dietary copper sources on the growth and intestinal microbial communities of Pacific white shrimp (Litopenaeus vannamei). Aquac. Nutr. 25: 828-840. Study 5

B. Effect of Copper Source on Shrimp Antioxidant Capacity and Immune Response

Key Findings Study Criteria • Partial or complete replacement shrimp fed Availa Cu at 0.5x rate of CuSO4 with Availa Cu at (Fig. 2c). This study evaluated 0.5x rate significantly improved effects of dietary copper • Activities of hepatopancreatic phenoloxidase activity in source on shrimp growth, Cu/Zn SOD, ALP and ACP were hemolymph (Fig. 2b). intestinal microbial highest when the treatment was communities, antioxidant • Partial or complete replacement of combined, but not statistically and immune response.

CuSO4 with Availa Cu at 0.5x rate different from Availa Cu at 0.5x. increased hepatopancreatic Cu/Zn • The inclusion of optimal levels SOD, ALP and ACP indicating better of Availa Cu in shrimp diets antioxidant capacity and immune improved shrimp health biomarkers, Treatment, ppm added to Control response in shrimp fed Availa Cu. demonstrating that robust shrimp Trace CuSO4 / AvCu Mineral Control CuSO4 AvCu 0.5x • Activity of hepatopancreatic LZM can be grown under commercial Copper 0 50 15/15 15

was significantly increased in conditions. a Diet contains 10 ppm of Copper coming from raw material.

Initial body weight: 1.86 g IBW Health Fig. 2 Stocking density: 30 shrimp/tank Feeding: 3x per day 2b. Hemolymph Antioxidant Capacity 2c. Hepatopancreas Antioxidant Replications: 3 and Immune Response Capacity and Immune Response Feeding period: 8 weeks Units/L Units/g protein Salinity: 25-28ppt

CuSO4 CuSO4/AvCu AvCu 0.5x

50000 35000 43% CP/8% Fat 40000 30000 ab b 30% fish meal, 3% krill 30000 25000 b ab meal, 22% soybean meal, a 20000 20000 6% poultry by-product, 150 b 80 b a 6% peanut meal b 60 b ab b 100 a b ab 40 a a a 50 a b ab a a a 20 0 0 Location: CuZn CP PO ALP ACP LZM CuZn ALP ACP LZM SOD SOD Ningbo University, China

Source: Yuan et al., 2019b. Effects of dietary dosage forms of copper supplementation on growth antioxidant capacity, innate immunity enzyme and gene expressions for juvenile Litopenaeus vannamei. Fish and Shellfish Immunology 84:1059-1067 Study 6

Effect of Zinc Source and Level on Shrimp Growth Performance, Meat Quality, Antioxidant and Immune Capacity

Key Findings

® • Availa Zn at 0.5x rate (60ppm Zn) of ZnSO4 • Availa Zn in combination with ZnSO4 or at maintained Specific Growth Rate (SGR) 0.5x rate translated to an economic advantage (Fig. 1a) and Feed Efficiency (FE) (Fig. 1b), showing over the control (ROI) of 45.6 % and 24.5%, Availa Zn is more effective than inorganic Zn. respectively.

• The combination of 60ppm Availa Zn + 60ppm • Best performance was with the combination of

ZnSO4 improved shrimp survival rate (data 60ppm Zn as ZnSO4 + 60ppm Zn as Availa Zn, not shown), SGR and FE by 4.3, 3.1 and 20% which indicates higher Zn supplementation may respectively (Fig. 1a & 1b). be required for maximized shrimp performance.

• Hepatopancreas antioxidant (Fig. 2a) and • ROI for Availa Zn at 0.5x rate is underestimated immune-related enzymes (Fig. 2b) were higher in due to the difficulty in attributing economic

shrimp fed a combination of ZnSO4 and Availa Zn value to benefits seen on antioxidant defense, or Availa Zn at 0.5x rate. immune response and product quality. Higher ROI is expected in the field and under more • Availa Zn in combination with ZnSO4 or at 0.5x challenging conditions. rate, reduced drip loss in muscle (Fig. 3a) and drip loss and thaw loss in whole shrimp (Fig. 3b).

Growth Performance Fig. 1

1a. Specific Growth Rate 1b. Feed Efficiency

3.55 0.8

3.45 z 0.6 z y y 3.35 0.4 x y y % per Day 3.25 0.2 x 3.15 0

Control ZnSO4 ZnSO4/AvZn AvZn 0.5x Control ZnSO4 ZnSO4/AvZn AvZn 0.5x

Source: Yuan et al., 2020. Aquaculture 522, 735120. Study Criteria

Health Fig. 2 This study aimed to provide novel insight into how the supplementation 2a. Hepatopancreas Cu-Zn SOD and level of a Zn source effected Units/mg protein whiteleg shrimp: growth response, 35 meat quality, oxidation resistance, 25 z z innate immunity . y 15 x Treatment, ppm added to Control

5 Trace ZnSO4 / AvZn Control ZnSO4 ZnSO4 /AvZn AvZn 0.5x Mineral Control ZnSO4 AvZn 0.5x Copper 0 120 60/60 60

2b. Hepatopancreas Enzyme Note: Basal diet contains 51 ppm of Zinc coming from raw material. Units/g protein

Control ZnSO4 ZnSO4 /AvZn AvZn 0.5x 35000 Initial body weight: 1.84g 30000 z Stocking density: 30 shrimp/tank 25000 20000 y Feeding: 3x per day xy 15000 Replications: 4 80 x Feeding period: 8 weeks 60 z z 40 y z Salinity: 25-28ppt x xy yz 20 x 0 AKP ACP LZM 43% CP/7.6% fat 30% fish meal, 3% krill meal, Meat Quality Fig. 3 22% soybean meal, 6% poultry by-product meal, 6% peanut meal

3a. Muscle Drip Loss, g/kg 3b. Whole Shrimp Drip Loss, g/kg Thaw Loss, g/kg Location: 90 80 Ningbo University, China

z z 80 60 z z y 70 y 40 z x y x 60 20 x w 50 0

Control ZnSO4 ZnSO4 / AvZn Control ZnSO4 ZnSO4 / AvZn AvZn 0.5x AvZn 0.5x Essential Trace Minerals for Shrimp

BENEFIT TRACE MINERALS DESCRIPTION

Disease Resistance Zinc, Manganese, • Humoral immunity Copper, Selenium, Iron • Cell-mediated immunity • Antioxidant activity to remove free radicals and protect cell membranes

Exoskeleton Health Zinc, Manganese, • Exoskeleton formation, development and reconstruction Copper during molting • Cell division and protein synthesis for normal tissue mineralization

Gut Integrity Zinc, Manganese, • Improves wound healing Copper • Epithelial tissue integrity through maintaining of cell division, protein synthesis and antioxidant activity to remove superoxide radicals

Muscle Development Zinc, Selenium, • Enzyme systems required for growth Chromium, Copper • Energy and protein metabolism • Cell membrane protection from peroxides • Influences carbohydrate, lipid and protein metabolism • Oxygen carrying function

Early Stage Zinc, Manganese, • Energy and protein metabolism Development Copper, Selenium • Cell proliferation • Normal tissue mineralization • Cell membrane protection • Hematopoiesis, the formation of hemocyanin, a large copper-containing protein that serve as the oxygen- carrying in the hemolymph

Meat Quality Zinc, Selenium, • Cell membrane protection Chromium, Iron • Antioxidant activity • Influences carbohydrate, lipid and protein metabolism • Enhanced meat color Shrimp Feeding Recommendations

Zinpro Recommendations Minimum Requirement Mineral ZPM Products ZPM, mg/kg diet

ZINPRO® Zn 60 Availa®Zn CuPlex® Cu 40 Availa®Cu MANPRO® Mn 40 Availa®Mn

Fe Availa®Fe 50

Ia 4

Seb Availa®Se 0.3

MiCroPlex® Crc 0.4 Availa®Cr

a Not a current ZPM source b Note upper limit allowed in EU is of 0.2 ppm, provided as organic source c Use where commercially available Zinpro exists to improveZinpro the wellnessexists to andimprove performance the wellness of animals and performance of animals and contribute to a healthier,and contribute more sustainable to a healthier, world. more sustainable world.

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