For the various benefits such as growth performance, reproduction, and enhancement of immunity, high dosage of zinc and copper are supplied to pigs. Commonly, 2000 to 2500 ppm of ZnO and 200 to 250 ppm of CuSO4 are used in pig industry. But those dosages of ZnO and CuSO4 is excess, then unabsorbed Z...
For the various benefits such as growth performance, reproduction, and enhancement of immunity, high dosage of zinc and copper are supplied to pigs. Commonly, 2000 to 2500 ppm of ZnO and 200 to 250 ppm of CuSO4 are used in pig industry. But those dosages of ZnO and CuSO4 is excess, then unabsorbed ZnO and CuSO4 are discharged as feces and that makes soil pollution by using as a compost. Moreover, there are some reports that high dose of ZnO and CuSo4 shows toxicity to tissues. It needs to minimize the usage of ZnO and CuSO4 as feed additives in pig feed with retaining positive effects of Zn and copper. Alternatively, we need to increase Zn and Copper utilization efficiency. In this study, we want to find out of strategy of zinc and copper supplementation that can replace current ZnO and CuSO4 additives. For this, we compare the effects of inorganic forms, ZnO and CuSO4 on intestinal immunity with their substitutes, which is better bioavailabilityin this study.
In the first experiment for the ZnO study, twenty piglets were used in a 2-week feeding trial to determine the effects of various dietary zinc forms of zinc. Glycine chelated ZnO and nanoparticle-size ZnO were used as substitutes that can absorb easily and have higher bioavailability. The dietary treatments included a negative control (CON), standard ZnO (ZnO, 2,500 ppm), glycine chelate zinc (Chelate-ZnO, 200 ppm), and nanoparticle-sized ZnO (Nano-ZnO, 200 ppm) for the first experiment. After 2 weeks experiment, mesenteric lymph node and lamina propria and the population of CD4+ T cell and subsets are analyzed by flow cytometry. Gene expression of major cytokines, tight junction protein, and antimicrobial peptide in Intestinal tissue and cell from mesenteric lymph node were measured by qRT-PCR.
For the secondary experiment, twenty-four weaned piglets were used in a 2-week feeding trial to determine the effects of differential ratio of zinc oxide and glycine chelate zinc oxide. T1 (Inorganic zinc : organic zinc = 850:150 ppm), T2 (Inorganic zinc : organic zinc = 700:300 ppm), T3 (Inorganic zinc : organic zinc = 500:500 ppm), and T4 (low protein diet + mixed additives [0.1% essential oil + 0.08% protease + 0.02% xylanase, MFA]) for the second experiment. After 2 weeks experiment, intestinal immune cells are isolated from mesenteric lymph node and lamina propria and the population of CD4+ T cell and subsets are analyzed by flow cytometry. The expression of genes for major cytokines and barrier function were measured by qRT-PCR.
In the first experiment, compared to the ZnO group, the Chelate-ZnO group exhibited higher proportion of Th1 and Treg cells and the Nano-ZnO group had higher numbers of Th2 and Th17 cells in the mesenteric lymph nodes. ZnO treatment increased IL-6 and IL-8 levels in the colon tissues and these positive effects were observed in Chleate-ZnO and Nano-ZnO with lower level. In second experiment, the proliferation of mesenteric lymph node cells was significantly increased after stimulation with LPS than non-stimulated cell in T3 (500:500) treated group. In the large intestinal lamina propria, Th1 and Th17 population were higher in T4 (LP+ mixed additives) than the other group that treated ZnO. Collectively, immunologically low dosage of glycine chelate ZnO can replace high dose of inorganic ZnO, also, the ratio of inorganic zinc oxide and glycine chelate zinc oxide had similar effect on intestinal immune cell.
For the study of CuSO4, thirty-two and thirty growing pigs were used in a 4-weeks feeding trial to find out immunologically appropriate substitutes and dosage. The substitutes that used in this study, Copper amino acid complex and copper glycine complex, have similar bioavailability and akin or higher result of growth performance. The control group (CON) fed with basal diet, while treatment groups were fed a basal diet supplemented with 100 mg/kg of copper sulfate (CuSO4), Cu-glycine complex (glycine), and Cu-amino acid complex (AA) for the first experiment. After feeding trial, the population of CD4+ T cell and subsets in mesenteric lymph node cell are analyzed by flow cytometry. Gene expression of major cytokines, tight junction protein, and antimicrobial peptide in Intestinal tissue and LPS stimulated cell from mesenteric lymph node were measured by qRT-PCR.
In second experiment, CON fed with basal diet, 50 ppm and 100 ppm of CuSO4 and copper-glycine complex were supplemented for 6-weeks. After feeding trial, intestinal immune cells are isolated from mesenteric lymph node and lamina propria and the population of CD4+ T cell and subsets are analyzed by flow cytometry. Gene expression of major cytokines, tight junction protein, and antimicrobial peptide in Intestinal were measured by qRT-PCR.
In the first experiment, T-bet+CD4+ T cell (Th1) and GATA3+CD4+ T cells (Th2) populations were significantly lower in AA treated group than the other groups in the small intestinal lamina propria. Also in the second experiment, 100 ppm of AA treated group had lower population of Th1 and Th2 cells. Also, 50 ppm of AA treatment decreased the population of Th1 cells but the population of Th2 cells were not changed in small intestine. Ultimately, 100 ppm of copper amino acid complex can replace the CuSO4 in pig additives.
In summary, immunologically low dosage of gylcine chelate zinc can replace high dosage of zinc oxide and copper amino acid complex can replace CuSO4 in pig fed additives. Consequently, excessive dosage zinc and copper supplementation can be reduced and that makes less environmental pollution by the feces of the pigs that contains high metal.
For the various benefits such as growth performance, reproduction, and enhancement of immunity, high dosage of zinc and copper are supplied to pigs. Commonly, 2000 to 2500 ppm of ZnO and 200 to 250 ppm of CuSO4 are used in pig industry. But those dosages of ZnO and CuSO4 is excess, then unabsorbed ZnO and CuSO4 are discharged as feces and that makes soil pollution by using as a compost. Moreover, there are some reports that high dose of ZnO and CuSo4 shows toxicity to tissues. It needs to minimize the usage of ZnO and CuSO4 as feed additives in pig feed with retaining positive effects of Zn and copper. Alternatively, we need to increase Zn and Copper utilization efficiency. In this study, we want to find out of strategy of zinc and copper supplementation that can replace current ZnO and CuSO4 additives. For this, we compare the effects of inorganic forms, ZnO and CuSO4 on intestinal immunity with their substitutes, which is better bioavailabilityin this study.
In the first experiment for the ZnO study, twenty piglets were used in a 2-week feeding trial to determine the effects of various dietary zinc forms of zinc. Glycine chelated ZnO and nanoparticle-size ZnO were used as substitutes that can absorb easily and have higher bioavailability. The dietary treatments included a negative control (CON), standard ZnO (ZnO, 2,500 ppm), glycine chelate zinc (Chelate-ZnO, 200 ppm), and nanoparticle-sized ZnO (Nano-ZnO, 200 ppm) for the first experiment. After 2 weeks experiment, mesenteric lymph node and lamina propria and the population of CD4+ T cell and subsets are analyzed by flow cytometry. Gene expression of major cytokines, tight junction protein, and antimicrobial peptide in Intestinal tissue and cell from mesenteric lymph node were measured by qRT-PCR.
For the secondary experiment, twenty-four weaned piglets were used in a 2-week feeding trial to determine the effects of differential ratio of zinc oxide and glycine chelate zinc oxide. T1 (Inorganic zinc : organic zinc = 850:150 ppm), T2 (Inorganic zinc : organic zinc = 700:300 ppm), T3 (Inorganic zinc : organic zinc = 500:500 ppm), and T4 (low protein diet + mixed additives [0.1% essential oil + 0.08% protease + 0.02% xylanase, MFA]) for the second experiment. After 2 weeks experiment, intestinal immune cells are isolated from mesenteric lymph node and lamina propria and the population of CD4+ T cell and subsets are analyzed by flow cytometry. The expression of genes for major cytokines and barrier function were measured by qRT-PCR.
In the first experiment, compared to the ZnO group, the Chelate-ZnO group exhibited higher proportion of Th1 and Treg cells and the Nano-ZnO group had higher numbers of Th2 and Th17 cells in the mesenteric lymph nodes. ZnO treatment increased IL-6 and IL-8 levels in the colon tissues and these positive effects were observed in Chleate-ZnO and Nano-ZnO with lower level. In second experiment, the proliferation of mesenteric lymph node cells was significantly increased after stimulation with LPS than non-stimulated cell in T3 (500:500) treated group. In the large intestinal lamina propria, Th1 and Th17 population were higher in T4 (LP+ mixed additives) than the other group that treated ZnO. Collectively, immunologically low dosage of glycine chelate ZnO can replace high dose of inorganic ZnO, also, the ratio of inorganic zinc oxide and glycine chelate zinc oxide had similar effect on intestinal immune cell.
For the study of CuSO4, thirty-two and thirty growing pigs were used in a 4-weeks feeding trial to find out immunologically appropriate substitutes and dosage. The substitutes that used in this study, Copper amino acid complex and copper glycine complex, have similar bioavailability and akin or higher result of growth performance. The control group (CON) fed with basal diet, while treatment groups were fed a basal diet supplemented with 100 mg/kg of copper sulfate (CuSO4), Cu-glycine complex (glycine), and Cu-amino acid complex (AA) for the first experiment. After feeding trial, the population of CD4+ T cell and subsets in mesenteric lymph node cell are analyzed by flow cytometry. Gene expression of major cytokines, tight junction protein, and antimicrobial peptide in Intestinal tissue and LPS stimulated cell from mesenteric lymph node were measured by qRT-PCR.
In second experiment, CON fed with basal diet, 50 ppm and 100 ppm of CuSO4 and copper-glycine complex were supplemented for 6-weeks. After feeding trial, intestinal immune cells are isolated from mesenteric lymph node and lamina propria and the population of CD4+ T cell and subsets are analyzed by flow cytometry. Gene expression of major cytokines, tight junction protein, and antimicrobial peptide in Intestinal were measured by qRT-PCR.
In the first experiment, T-bet+CD4+ T cell (Th1) and GATA3+CD4+ T cells (Th2) populations were significantly lower in AA treated group than the other groups in the small intestinal lamina propria. Also in the second experiment, 100 ppm of AA treated group had lower population of Th1 and Th2 cells. Also, 50 ppm of AA treatment decreased the population of Th1 cells but the population of Th2 cells were not changed in small intestine. Ultimately, 100 ppm of copper amino acid complex can replace the CuSO4 in pig additives.
In summary, immunologically low dosage of gylcine chelate zinc can replace high dosage of zinc oxide and copper amino acid complex can replace CuSO4 in pig fed additives. Consequently, excessive dosage zinc and copper supplementation can be reduced and that makes less environmental pollution by the feces of the pigs that contains high metal.
주제어
#pig intestine immune zinc copper
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