Z Enterosorbent on Serum Reproductive Hormone, Immunoglobulin, Antibody Titer in Young Pigs Fed Purified Zearalenone

AMELIORATE EFFECT OF CALIBRIN®-Z ENTEROSORBENT ON SERUM REPRODUCTIVE HORMONE, IMMUNOGLOBULIN, ANTIBODY TITER IN YOUNG PIGS FED PURIFIED ZEARALENONE

Ron Cravens1, S. Z. Jiang2, Z. B. Yang2, Ron Cravens, Amlan International, Inc, Chicago, IL, US1, phone (312) 321-1515 [email protected]; Department of Animal Sciences W. R. Yang2, Sara Johnston1, Fang Chi1 and Technology, Shandong Agricultural University, Taian, Shandong, China2

ABSTRACT TABLE 1. Antibody Titer to Classic Swine Fever of Pigs Fed Zearalenone Contaminated Diets Zearalenone (ZEA) is a mycotoxin that mimics estrogen; causing infertility, abortion, With or Without Calibrin-Z and other reproductive problems, especially in swine. Besides its estrogenic effects, Day From Vaccination Control Control + ZEA ZEA + ZEA + ZEA + P values CZ1 1 ppm CZ1 CZ2 CZ4 the toxin can also negatively impact the immune response. Calibrin-Z (CAZ), a -1 4.50 4.83 4.67 4.33 4.67 4.50 0.980 highly-refined montmorillonite sorbent mineral, has high affinity and capacity to 7 4.83 5.17 4.33 4.50 4.83 4.83 0.787 14 5.50 5.67 5.33 5.33 5.50 5.50 0.127 sequester a wide range of mycotoxins found in feed grains. The objective of this study 21 6.83a 7.33a 6.00b 6.00b 6.17b 6.00b 0.009 was to evaluate different dosages of CAZ on the effects of ZEA on serum hormone and immune response in piglets. A total of 36 pigs with initial body weight of 8.9 ± TABLE 2. Serum Markers of pigs Fed Zearalenone Contaminated Diets With or Without Calibrin-Z

0.2 kg were allotted to 6 treatments (TRT). The 6 TRT were: 1) Control (Con); 2) Con + Parameter Control Control + ZEA ZEA + ZEA + ZEA + CZ1 1 ppm CZ1 CZ2 CZ4 0.1% CAZ; 3) Con + 1 ppm ZEA; 4) Con + 1 ppm ZEA + 0.1% CAZ; 5) Con + 1ppm Aspartate aminotransferase (U/L) 68b 66b 86a 80a 76ab 71ab ZEA + 0.2% CAZ; 6) Con + 1 ppm ZEA + 0.4% CAZ. Data were analyzed using the Alanine aminotransferase (U/L) 58b 56b 78a 73a 68ab 61b Alkaline phosphatase (U/L) 201b 197b 246a 240a 231a 220ab GLM procedure of SAS with individual pig as the basis for analysis. Each pig received γ-Glutamyltransferase (U/L) 23b 27b 41a 37a 36a 31ab classical swine fever (CSF) vaccine 1-d before the study, serum samples were Urea (mmol/L) 2.7b 2.9b 4.1a 3.7a 3.4ab 3.0b Creatinine (µmol/L) 77b 75b 88a 91a 79ab 75b collected weekly for CSF antibody titer determination. On d 21, blood and spleen samples were collected for hormone, immunoglobulin, interleukin (IL) cytokine FIGURE 1. Serum estrodial of pigs fed zearalenone contaminated diets with analyses, and lymphocyte proliferation rate (LPR). Gilts fed TRT 3 had lower (P < or without Calibrin-Z

0.05) progesterone (75% of 1 and 2), and testosterone (74% of 1 and 2) levels than 30.00 TRT 1 and 2. Adding CAZ improved serum hormone levels and the elevated hormone a a Control was CAZ dosage dependent. Male pigs fed TRT 3 showed similar results as gilts 24.00 b Control + CZ1 a a a c ZEA 1 ppm b c except serum estrodial was not different. Serum IgA and IgM were not different; 18.00 c c c ZEA + CZ1 a c a however, IgG level was reduced (P < 0.01) in TRT 3 (77% of 1 and 2) compared with a b TRT 1 and 2. It increased linearly (P < 0.05) in TRT 4, 5, and 6 but remained lower (P pg/mL 12.00 ZEA + CZ2 ZEA + CZ4 < 0.05) than TRT 1 and 2 (84% of 1 and 2). The LPR from blood and spleen cells 6.00 followed a trend similar to IgG. Serum IL-2 followed results similar to those of the 0.00 hormones; pigs fed TRT 3 had the lowest IL-2 but an addition of 0.4% clay restored Female Male the levels equal to TRT 1 and 2. There was no difference on d-7 or d-14 on CFS titers. FIGURE 2. Serum progesterone of pigs fed zearalenone contaminated diets with On d-21, pigs fed TRT 2 had the highest titer against CSF and greater (P < 0.01) than or without Calibrin-Z pigs fed TRT 3, 4, 5, and 6; but not different than TRT 1. Feeding 1 ppm of purified 0.50 ZEA caused adverse effects on immunity in pigs; which were ameliorated by CAZ. a a Control ab ab Feeding CAZ without ZEA had the greatest antibody titer production, implying that the 0.40 b Control + CZ1 b a ZEA 1 ppm Calibrin-Z may add value to feed with or without a mycotoxin challenge. a a c c 0.30 ab ZEA + CZ1 b b ng/mL a ZEA + CZ2 OBJECTIVE 0.20 ab b ab The objective of this study was to evaluate the ameliorative effect of Calibrin-Z on the effects ZEA + CZ4 of zearalenone (ZEA) serum reproductive hormone and immune response in young pigs. 0.10 c c 0.00 MATERIALS & METHODS Female Male FIGURE 3. Serum testosterone of pigs fed zearalenone contaminated diets 36 weaning pigs (1/2 male, 1/2 female) Body Weight 8.9 ± 0.2kg with or without Calibrin-Z

100.00 a a Control Treatments ab c ab c a b Control + CZ1 • Control 80.00 a a b a ZEA 1 ppm • Control + 0.1% Calibrin-Z (CZ1) b 60.00 b • Control + 1 ppm ZEA ZEA + CZ1

• Control + 1 ppm ZEA + 0.1% Calibrin-Z (CZ1) ng/mL 40.00 ZEA + CZ2 • Control + 1 ppm ZEA + 0.2% Calibrin-Z (CZ2) ZEA + CZ4 • Control + 1 ppm ZEA + 0.4% Calibrin-Z (CZ4) 20.00 a ab ab c bc ab 0.00 The feeding period was 21 d. Each pig received classical swine fever (CSF) vaccine 1 d Female Male before the study, serum samples were collected weekly for CSF antibody titer FIGURE 4. CD4 and CD8 T-cell counts of pigs fed zearalenone contaminated diets determination. On d 21, blood and spleen samples were collected for hormone, with or without Calibrin-Z immunoglobulin, interleukin (IL) cytokine analyses, and lymphocyte proliferation rate 50.00 a a Control a c ab a b c (LPR). Calibrin-Z is calcium montmorillonite clay that has been further processed, ab ab ab Control + CZ1 40.00 b b resulting in increased binding of ZEA and other mycotoxins. b lear Cells) ZEA 1 ppm a 30.00 a ab RESULTS b b ab ZEA + CZ1 20.00 ZEA + CZ2 Adding Calibrin-Z (CAZ) improved serum hormone levels and the elevated hormone ZEA + CZ4 eripheral Blood Mononu c was CAZ dosage dependent. Male pigs fed TRT 3 showed similar results as gilts ( P 10.00 except serum estrodial was not different. Serum IgA and IgM were not different, IgG % 0.00 level was reduced (P < 0.01) in TRT 3 (77% of 1 and 2) compared with TRT 1 and 2. It CD4 CD8 increased linearly (P < 0.05) in TRT 4, 5, and 6 but remained lower (P < 0.05) than TRT 1 and 2 (84% of 1 and 2).The LPR from blood and spleen cells followed a trend CONCLUSIONS similar to IgG. Serum IL-2 followed results similar to those of the hormones; pigs fed • Feeding 1 ppm of puriﬁed ZEA caused adverse effects on immunity TRT 3 had the lowest IL-2 but an addition of 0.4% CAZ restored the levels equal to in pigs; which were ameliorated by Calibrin-Z. TRT 1 and 2. There was no difference on d-7 or d-14 on CFS titers. On d-21, pigs fed TRT 2 had the highest titer against CSF and greater (P < 0.01) than pigs fed TRT 3, 4, • Feeding Calibrin-Z without ZEA had the greatest antibody titer 5, and 6; but not different than TRT 1. production, implying that the Calibrin-Z adds value to feed with or without a mycotoxin challenge.