Supplementation of Zinc on Antioxidant Activity, Blood Profile, Mineral Availability, Abdominal Fat, Digestive and Accessory Organs of Sikumbang Janti Duck

1. Introduction

Zinc (Zn) is a mineral (micro) that is important for the growth and development of poultry, especially in the process of enzymes catalysis, structural, and regulatory ( 1 , 2 ). In recent years, research related to Zn in poultry has been expanding. Zinc addition in poultry feed can improve performance ( 3 ), egg quality ( 4 ), antioxidant activity ( 3 , 5 ), immune system ( 5 - 7 ) and reduce stress caused by increasing environmental temperature ( 8 ).

Zinc minerals are necessary for tropical nations such as Indonesia, where the average daily temperature exceeds the comfort zone of poultry, including laying ducks. The Sikumbang Janti duck is one of the local egg-laying ducks that has the potential to be developed in the Indonesian state of West Sumatra. The Sikumbang Janti duck is a laying duck from Payakumbuh City with a dominant white coat color, except for green primary wing feathers and a black beak ( 9 - 11 ). However, Sikumbang Janti duck egg production is still relatively low, with an average production of 190-210 eggs/year ( 10 ). Therefore, supplementation of the mineral Zn in feed is expected to increase production. Apart from functioning as an antioxidant, the mineral Zn plays an essential role in regulating the hormone system, cell growth, immunity, and reproduction ( 12 ).

Previous research has demonstrated that zinc increases the egg production of Magelang ducks ( 13 ). It was also mentioned that the Zn mineral could overcome the effects of heat stress by reducing oxidative damage to cell membranes caused by free radicals and reducing the heterophil/lymphocyte ratio and the mortality rate of laying duck embryos ( 14 ). Nevertheless, there is no data on zinc addition to the diet of Sikumbang Janti ducks, especially antioxidant activity, blood profile, mineral availability, abdominal fat, etc. Therefore, the present study aimed to evaluate the effect of zinc addition on antioxidant activity, blood profile, mineral availability, and abdominal fat of Sikumbang Janti duck.

2. Materıals and Methods

2.1. Animal, design, and diet

This study used 96 female Sikumbang Janti ducks aged eight weeks and weighing an average of 752.41 ± 81.99 g/bird. The Sikumbang Janti duck was purchased from a farmer from Payakumbuh City, West Sumatra. The current research lasted eight weeks using a completely randomized design with four treatments and four replicates Z0 (control diet); Z1 (Z0 + 30 mg Zn/kg), Z2 (Z0 + 60 mg Zn/kg), and Z3 (Z0 + 90 mg Zn/kg) (6 birds per replicate). The cages used in this study were 1.5m x 1.5m. The cage provided a place to eat and drink. The Zn used was 75% ZnO (Zn-O-India). The ration (Table 1) was formulated with an energy metabolism of 2,700 kcal/kg and 17% crude protein ( 15 ).

2.2. Parameters Observed

2.2.1. Antioxidant Activity

At the end of the study, one duck per repetition was slaughtered, and thigh meat was taken to analyse antioxidant activity. Antioxidant activity was performed with the 1.1-diphenyl-2picrylhydrazil (DPPH) method ( 16 ).). The specimen was dissolved (1 mg ml-1) in methanol. The mixture was vortexed after adding 500 μl of DPPH (125 μM in methanol) to 500 μl of the sample in a test tube. After 30 min at room temperature, the absorbance of the solution was measured at a wavelength of 517 nm using a spectrophotometer (Shimadzu UV VIS -1800, Japan). The IC₅₀ value was determined using the linear regression equation, which describes the relationship between sample concentration (test compound) with Symbol X and average radical scavenging activity (measurement replication series, symbol Y). The higher the antioxidant activity, the lower the IC₅₀ value ( 17 ).

$\mathrm{DPPH\; radical\; scavenging\; activity\; \%}=\frac{\mathrm{A\; Sample}-\mathrm{A\; Control}}{\mathrm{A\; Control}}\times 100$

2.2.2. Blood Profile

Blood sampling was carried out at the end of rearing (1 duck per replication) in the brachial vein using a sterile syringe. Blood samples were put into tubes containing ethylene diamine tetra acetic acid (EDTA). Blood analysis consisted of erythrocytes, leukocytes, heterophiles, lymphocytes, and monocytes using a hemocytometer and microscope (Olympus CX 23, China), hemoglobin using a complete reagent kit (Merckotest^®) and hematocrit using Micro-Capillary reader (USA).

2.2.3. Mineral Availability of Tibia

The tibia of each duck was cleaned, and muscle and attached cartilage were removed, washed with water, and oven-dried (Memmert, Germany) at 60ºC for 24 h. The procedure for analysing the minerals Calcium (Ca), phosphor (P), and Zn followed the analysis method of association of official analytical chemist ( 18 ).

2.2.4. Abdominal Fat

Abdominal fat following the method outlined by Reski et al. ( 19 ). Sikumbang Janti ducks were slaughtered (one duck per repetition) at the end of the experiment (16 weeks). A digital balance (Osaka-HWH®, Japan) was used to test parameters.

2.3. Data analysis

An analysis of variance was performed on the data based on a 4 x 4 complete randomized design, and Duncan's multiple range test was performed on the means. A P < 0.05 was considered statistically significant.

3. Results and Discussion

3.1. Antioxidant Activity

The average antioxidant activity and Zn concentration in Sikumbang Janti duck meat are presented in Table 2. Zn addition on feed significantly increased antioxidant activity (DPPH) compared to the treatment without Zn (P<0.01). Zinc concentration in thigh meat was significantly affected by Zn feed (P<0.01). Adding up to 90 mg Zn/kg of feed enhanced the Zn concentration in thigh meat compared to treatment without Zn. However, the Zn addition of 90 mg Zn/kg did not affect Zn concentration in thigh meat compared to the Zn addition of 30 mg Zn/kg feed.

Zinc addition in feed has a positive effect on antioxidant status. In this study, the measurement of antioxidant activity using the DPPH method showed that Zn has antioxidant properties with a mechanism as a radical scavenger. The higher the Zn addition in the Sikumbang Janti duck ration, the higher the Zn deposited in the meat, which has an impact on the higher antioxidant activity. The lower the value of the antioxidant activity (IC₅₀) reflects, the higher the antioxidant activity and vice versa ( 17 ).

Some research results also showed that Zn addition in duck feed ( 2 , 20 ), broiler ( 3 ), laying hens ( 5 , 21 ), and quail ( 22 ) can improve or enhance antioxidant status. Zinc prevents oxidation of the sulfhydryl group and reduces the generation of reactive oxygen species. Zinc is a cofactor for the enzyme Cu Zn-superoxide dismutase, which inhibits free radicals by facilitating the conversion of superoxide anions to hydrogen peroxide ( 21 ). In addition, Zn increases the formation of metallothionein, which serves as a hydroxyl radical scavenger ( 23 ).

3.2. Blood Profile

Table 3 presents the average blood profile of Sikumbang Janti ducks. Leukocytes increased (P<0.05) when Zn feed addition (60 mg Zn/kg) was compared to without Zn. Zinc addition did not affect erythrocytes, hemoglobin, hematocrit, heterophils, lymphocytes, monocytes, or the H:L ratio (P>0.05).

Animal blood profile describes the nutritional and physiological status according to the internal and external environment. In general, the blood profile of poultry is influenced by physiological (age, sex, reproductive status and season) and pathological (disease) factors ( 24 ). In this study, Zn addition up to 90 mg Zn/kg in the diet did not affect blood profile (except leukocytes) with average range values. This shows that the Sikumbang Janti ducks were in normal physiological conditions during the study. In this study, Z2 treatment resulted in the highest number of leukocytes compared to Z0 treatment. Zinc has a relationship with the formation of leukocytes, and this is because Zn has a role in Thymulin hormone activity. Thymulin is secreted by thymic epithelial cells, which plays a role in the maturation of T lymphocytes, where Zn deficiency can cause apoptosis in lymphoid cells, which results in a reduction in total leukocytes and lymphocytes ( 25 ).

3.3. Mineral Availability of Tibia

Table 4 shows the average availability of the minerals Ca, P, and Zn in the tibia. Zinc addition significantly affected Ca, P, and Zn content in the tibia (P<0.01). The addition of 60 mg Zn/kg resulted in higher Ca, P, and Zn content compared to the 30 mg Zn/kg treatment and the treatment without Zn. However, addition at the highest level (90 mg Zn/kg) showed a decrease in the P and Zn content of the tibia (P<0.01).

Bone is a complex tissue that supports muscle, body growth and development and is extremely important as a reserve of calcium and phosphorus in the formation of eggshells ( 26 ). This study shows the content of minerals (Ca, P, and Zn) in the tibia bone is affected by Zn addition in the diet. Zinc has an essential role in bone calcification ( 27 ). Zinc addition up to 60 mg Zn/kg (Z2 treatment) in the ration increases the content of Ca, P, and Zn in the tibia. Some research indicates that supplementing broiler diets with 80 ppm of zinc leads to a calcium content of up to 51.79% ( 29 ). A high dose (80 ppm) of zinc addition resulted in a more significant zinc content in the tibia than a low dose (20 ppm) ( 30 ). Zinc in bone is a functional Zn reserve that can be easily mobilized for tissue needs. Zinc addition up to 48 mg Zn/kg laying hens had higher tibial bone strength and calcium deposition than other treatments ( 5 ).

In addition, Zn influences the average physiological effect of vitamin D on calcium metabolism, influencing calcium deposition in bone tissue ( 31 ). In this study, addition at high levels (90 mg Zn/kg) showed a decrease in tibial bone phosphorus concentrations, indicating that excessive Zn levels harm the bioavailability of other minerals. The antagonistic nature of Zn and phosphorus minerals may explain this phenomenon ( 23 ).

3.4. Abdominal Fat

Figure 1 shows the average abdominal fat of the Sikumbang Janti duck. Zinc addition at 60 mg/kg significantly (P<0.05) reduced abdominal fat compared to Z0 and Z1 treatment. However, it was no different from the Z3 group (90 mg/kg).

Figure 1. Abdominal fat of Sikumbang Janti Duck fed by a zinc-contained diet. Means in the same bar chart with different superscripts differ significantly (P<0.01)

The weight of abdominal fat in this study decreased as Zn addition of the diet was raised (Figure 1). According to Mohammadi ( 32 ), supplementing the diet with zinc (Zn-SO₄ and Zn-Met) lowered the proportion of abdominal fat compared to the control treatment. In different studies, the lack of an increase in abdominal fat could be accounted for by differences in species and Zn content in the diet ( 33 ).

These findings are consistent with previous research indicating that organic and inorganic zinc addition in the feed had no negative impact on the visceral and digestive organs of Pekin ducks.

Addition of dietary Zn at 60 mg Zn/kg in diet increases antioxidant activity, leukocytes, zinc concentration in thigh meat, mineral availability in the tibia, and decreases abdominal fat weight of Sikumbang Janti duck.

Authors' Contribution

Study concept and design: R.K.R, Acquisition of data: R.K.R; R. A, Analysis and interpretation of data: R.K.R, Drafting of the manuscript: R.K.R; A.D; Z, Critical revision of the manuscript for important intellectual content: K.S; K, Statistical analysis: R. A, Administrative, technical, and material support: S. R

Ethis

All research methods were approved by The Research Ethics Committee Faculty of Medicine Universitas Andalas, number: 31/UN.16.2/KEP-FK/2023.

Conflict of Interest

The authors declares that there is no conflict of interest with any financial, personal, or other relationships with other people or organization related to the material discussed in the manuscript.

References

1. Sahin K, Sahin N, Kucuk O, Hayirli A, Prasad AS. Role of dietary zinc in heat-stressed poultry: A review. Vol. 88, Poultry Science. Poultry Science Association; 2009.
2. Zhang YN, Wang S, Li KC, Ruan D, Chen W, Xia WG, et al. Estimation of dietary zinc requirement for laying duck breeders: effects on productive and reproductive performance, egg quality, tibial characteristics, plasma biochemical and antioxidant indices, and zinc deposition. Poult Sci. 2020; 99(1):454-62.
3. Hidayat C, Sumiati S, Jayanegara A, Wina E. Supplementation of Dietary Nano Zn-Phytogenic on Performance, Antioxidant Activity, and Population of Intestinal Pathogenic Bacteria in Broiler Chickens. Tropical Animal Science Journal. 2021; 44(1):90-9.
4. Al-Obaidi BMW, Mahmood EK, Alnoori MA, Alnori HM, Saeed OA. Effect of organic zinc supplementation into basal diets on productive performance of laying hens. J Indones Trop Anim Agric [Internet]. 2022;47(4):257-64. Available from: https://ejournal.undip.ac.id/index.php/jitaa/article/view/46518.
5. Li LL, Gong YJ, Zhan HQ, Zheng YX, Zou XT. Effects of dietary Zn-methionine supplementation on the laying performance, egg quality, antioxidant capacity, and serum parameters of laying hens. Poult Sci. 2019; 98(2):923-31.
6. Sunder GS, Panda AK, Gopinath NCS, Rama Rao S v., Raju MVLN, Reddy MR, et al. Effects of higher levels of zinc supplementation on performance, mineral availability, and immune competence in broiler chickens. Journal of Applied Poultry Research. 2008; 17(1):79-86.
7. Moghaddam HN, Jahanian R. 2009_AJAS_Moghaddam - Zn, Immune. Asian-Aust J Anim Sci. 2009; 22(3):396-403.
8. Ramiah SK, Awad EA, Mookiah S, Idrus Z. Effects of zinc oxide nanoparticles on growth performance and concentrations of malondialdehyde, zinc in tissues, and corticosterone in broiler chickens under heat stress conditions. Poult Sci. 2019; 98(9):3828-38.
9. Arlina F, Husmaini S, Rhoudha R, Sardi WR, Rafian T. Keragaman Fenotipe Kualitatif dan Kuantitatif Itik Sikumbang Jonti sebagai Plasma Nutfah di Sumatera Barat. Jurnal Ilmu Peternakan dan Veteriner Tropis (Journal of Tropical Animal and Veterinary Science).. 2022; 11(3):291.
10. Nova TD, Anggraeni SD, Wardiansyah M, Ramadhani EP. Frekuensi Pemberian Ransum Secara Periodik dan Level Protein Terhadap Karkas dan Gambaran Darah Itik Lokal Sikumbang Janti. Jurnal Peternakan Indonesia (Indonesian Journal of Animal Science).. 2019; 21(2):64.
11. Rafian T, Yurnalis Y, Fenita Y, Iskandarsyah R. Polymorphism of Prolactin Gene (PRL/PstI) In Sikumbang Jonti Duck Using PCR-RFLP Methods. Jurnal Sain Peternakan Indonesia. 2022; 17(3):170-4.
12. Nasiadek M, Stragierowicz J, Klimczak M, Kilanowicz A. The role of zinc in selected female reproductive system disorders. Vol. 12, Nutrients. MDPI AG; 2020.
13. Darmawan A, Wiryawan KG, Sumiati S. Egg production and quality of magelang duck fed diets containing different ratio of omega 3: Omega 6 and organic zn. Media Peternakan Fakultas Peternakan Institut Pertanian Bogor. 2013; 36(3):197-202.
14. awad awad, el-shhat abdel ghany, abdelmaged marwa, shazly S, abdelhaleem hassan, ragab mona. Productıve And Reproductıve Performance Of Local Domyatı Ducks Fed Dıet Enrıched Wıth Organıc Zınc Durıng Summer Season. Egyptian Poultry Science Journal [Internet]. 2022 Sep 1 [cited 2023 Jan 22];42(3):313-30. Available from: https://epsj.journals.ekb.eg/article_264654.html.
15. Sinurat AP. Penyusunan ransum ayam buras dan itik. Dinas Peternakan DKI Jakarta: Jakarta; 2000.
16. Waksman De Torres N, Salazar-Aranda R, Pérez-López LA, López-Arroyo J, Alanís-Garza BA. Antimicrobial and antioxidant activities of plants from northeast of Mexico. Evidence-based Complementary and Alternative Medicine. 2011; 2011
17. Sánchez-Moreno C, A. Larrauri J, Saura-Calixto F. Free radical scavenging capacity and inhibition of lipid oxidation of wines, grape juices and related polyphenolic constituents. Food Research International. 1999; 32(6):407-12.
18. Association of Official Agricultural Chemists. Official Methods of Analysis [Internet]. 21st Edition. Rockville, MD 20850, United States of America: AOAC INTERNATIONAL; 2019 [cited 2023 Jan 22]. Available from: https://www.aoac.org/official-methods-of-analysis-21st-edition-2019/.
19. Reski S, Mahata ME, Rusli RK. The Impact of Dietary Fermented Seaweed (Turbinaria murayana) with Fruit Indigenous Micro Organism’s (IMO’s) as a Starter on Broiler Performance, Carcass Yield and Giblet Percentage. Adv Anim Vet Sci. 2022; 10(7):1451-7.
20. Wu XP, Zhu YF, Zhang KY, Ding XM, Bai SP, Wang JP, et al. Growth performance, zinc tissue content, and intestinal health in meat ducks fed different specific surface area of micronized zinc oxide. Poult Sci. 2019; 98(9):3894-901.
21. Yu Q, Liu H, Yang K, Tang X, Chen S, Ajuwon KM, et al. Effect of the level and source of supplementary dietary zinc on egg production, quality, and zinc content and on serum antioxidant parameters and zinc concentration in laying hens. Poult Sci. 2020; 99(11):6233-8.
22. Reda FM, El-Saadony MT, El-Rayes TK, Attia AI, El-Sayed SAA, Ahmed SYA, et al. Use of biological nano zinc as a feed additive in quail nutrition: biosynthesis, antimicrobial activity and its effect on growth, feed utilisation, blood metabolites and intestinal microbiota. Ital J Anim Sci. 2021; 20(1):324-35.
23. Niknia AD, Vakili R, Tahmasbi AM. Zinc supplementation improves antioxidant status, and organic zinc is more efficient than inorganic zinc in improving the bone strength of aged laying hens. Vet Med Sci. 2022.
24. Clark P, Boardman W, Raidal S. Atlas of Clinical Avian Hematology. Wiley Blackwell: New Jersey; 2009.
25. Omeya YS, Anihata JT, Ato SS, Awano FK, Hirato KS, Ugiyama MS, et al. Zinc-Deficiency Induced Changes in the Distribution of Rat White Blood Cells. Vol. 55, J Nutr Sci Vitaminol. 2009.
26. Bao YM, Choct M, Iji PA, Bruerton K. Effect of Organically Complexed Copper, Iron, Manganese, and Zinc on Broiler Performance, Mineral Excretion, and Accumulation in Tissues [Internet]. 2007. Available from: http://japr.oxfordjournals.org/.
27. Huang YL, Lu L, Xie JJ, Li SF, Li XL, Liu SB, et al. Relative bioavailabilities of organic zinc sources with different chelation strengths for broilers fed diets with low or high phytate content. Anim Feed Sci Technol. 2013; 179(1-4):144-8.
28. Star L, van der Klis JD, Rapp C, Ward TL. Bioavailability of organic and inorganic zinc sources in male broilers. Poult Sci. 2012; 91(12):3115-20.
29. Rayani TF, Mutia R, Sumiati S. Supplementation of zinc and vitamin E on apparent digestibility of nutrient, carcass traits, and mineral availability in broiler chickens. Media Peternakan Fakultas Peternakan Institut Pertanian Bogor. 2017; 40(1):20-7.
30. Olukosi OA, van Kuijk S, Han Y. Copper and zinc sources and levels of zinc inclusion influence growth performance, tissue trace mineral content, and carcass yield of broiler chickens. Poult Sci. 2018; 97(11):3891-8.
31. Mahdavi-Roshan M, Ebrahimi M, Ebrahimi A. Copper, magnesium, zinc and calcium status in osteopenic and osteoporotic post-menopausal women. Vol. 12, Clinical Cases in Mineral and Bone Metabolism. 2015.
32. Mohammadi V, Ghazanfari S, Mohammadi-Sangcheshmeh A, Nazaran MH. Comparative effects of zinc-nano complexes, zinc-sulphate and zinc-methionine on performance in broiler chickens. Br Poult Sci. 2015; 56(4):486-93.
33. Hidayat C, Sumiati S, Jayanegara A, Wina E. Effect of zinc on the immune response and production performance of broilers: A meta-analysis. Asian-Australas J Anim Sci. 2020; 33(3):465-79.
34. Attia YA, Abd Al-Hamid AE, Zeweil HS, Qota EM, Bovera F, Monastra G, et al. Effect of dietary amounts of inorganic and organic zinc on productive and physiological traits of White Pekin ducks. Animal. 2013; 7(6):895-900.