Effect of Prenatal Trace Mineral Source on Neonatal and Growing Calf Liver and Serum Mineral Status

Effect of Prenatal Trace Mineral Source on Neonatal and Growing Calf Liver and Serum Mineral Status D. M. Price* 1, A. F. Swain 1, J. M. Guevera 2, C. R. Trcalek 2, M. M. O'Neil 1, M. Irsik 2, O. Rae 2, M. J. Hersom 1, J. V. Yelich 1 ; Synopsis Calf trace mineral status varies across days after calving and is influenced by trace mineral source and breed. In this particular year, mineral source did affect calf performance. Summary The effect of cow prenatal trace mineral (TM) supplement source on calf TM status from birth through 3, 115 days of age, and weaning was examined. Factorial treatment (TRT) arrangements (Angus=, n=95 and Brangus=, n=96 cows; Inorganic=ING, n=98, and Organic=ORG yeast, n=93) utilized calves born to cows supplemented (3 day/week at a rate of 1 lb 1 1,000 lb BW 1 day in a pellet) with TM 90 d before expected parturition. Calf BW were collected at birth (n=191). A subset of calves (n=43) had bodyweight, serum and plasma collected by jugular vein puncture at birth (0 hour, before colostrum intake), 12 hour, 24 hour, 30, 115 days of age, and weaning. A subset of calves (ING, n=12, 6/breed, ORG, n=14, 7/breed) had liver biopsies performed and serum collected by jugular vein puncture on day 115 and weaning. Processed samples were frozen at 20 C and plasma was refrigerated until analyzed for TM (serum: Co, Cu, Fe, Mn, Mo, Se, Zn; plasma: Se). Time affected (P<0.02) all 0-24 hour serum and plasma TM except for Co (P=0.60). Calf Mn concentrations were undetectable at 0-24 hour. Copper status was deficient, whereas other minerals were marginal to adequate at the 0-24 hour period. On day 30, Se concentrations (whole blood and serum) were greater (P<0.05) for ORG than ING, Co concentration was greater for ING compared to ORG. At weaning, ORG had greater (P 0.02) body weight and ADG than ING. Serum Co and Zn were affected (P<0.001) by day, while all liver TM except for Co were affected (P<0.03) by day. The ORG calves had greater (P 0.01) serum Se, liver Se and Mn than ING. The calves had greater (P 0.04) serum Co, Se, and liver Co, Fe, and Zn, while had greater (P<0.05) serum Mo and liver Mn. These data demonstrate calf TM status varies by time, prenatal TM source and breed. Introduction Recent research has shown that altering the environment the fetus is exposed to during gestation can have significant effects on the performance of that animal in its postnatal life, a concept termed fetal programming. This originally started with the nutrient restriction model, which resulted in increased adipose tissue deposition as well as decreased muscle development (Zhu et al., 2006; Du et al., 2010) leading to alterations in carcass composition (Larson et al., 2009; Underwood et al., 2010). Trace minerals have traditionally been supplemented to cattle diets as inorganic salts. In spite of this tradition, recent attention has been placed on the use of organic or chelated trace mineral supplementation in the rumen diets. Organic trace minerals differ from inorganic forms as a result of their chemical association with an organic ligand. Numerous groups of these organic trace minerals are formed from this mineral-organic ligand combination, which are available in the animal feeding industry and include chelates, proteinates, and complexes (AAFCO, 2000). The recent attention towards organic mineral 1 Department of Animal Sciences, University of Florida, Gainesville, FL 2 College of Veterinary Medicine, University of Florida, Gainesville, FL

supplementation in ruminant diets has been fueled by numerous studies that have shown an increase in overall performance in cattle supplemented with organic minerals. It is generally accepted that this association is due to the increased bioavailability of organic minerals. Minerals play a significant role in many metabolic processes that affect growth performance, reproductive efficiency and immune function. Selecting the correct mineral supplement is crucial for maintaining these processes. With research clearly demonstrating across animal species that organic minerals are move bioavailable to the animal further investigation is imperative to determine whether the greater bioavailability of these organic mineral will have a more positive effect on the overall performance of beef cattle reproduction. Materials and Methods Pregnant Angus (, n=95) and Brangus (, n=96) cows were allotted to two mineral supplements containing either inorganic (ING, n=98) or organic (ORG, n=93) trace minerals in a two by two factorial design. Mineral supplementation began approximately 82±2 days for all cows (range 35 to 147 days) and 72±1 day for cows whose calves had blood and liver biopsies collected for trace mineral status prior to calving and continued through weaning. From the initiation of the experiment to the end of the breeding season cows grazed on dormant bahiagrass pastures, fed large round bales of hay for ad libitum consumption, and supplemented with soybean hulls to maintain a body condition score of 5 on a 9 point scale. Cows received the trace mineral supplement in a pelleted supplement fed at a rate of 1 lb per 100 lbs of cow body weight. Trace mineral and soybean hull supplements were offered three days per week. After the breeding season, cows grazed bahiagrass pastures in large mineral treatment groups. Trace mineral supplement was provided as a loose mineral for consumption based on formulated guidelines. Mineral was distributed on a weekly basis plus 10% to ensure appropriate consumption. The ING trace mineral supplement was formulated to meet the mature beef cow mineral requirements based on NRC recommendations (Table 1). The ORG trace mineral supplement was formulated to meet beef cow NRC requirements based on the assumption of greater bioavailability for organic sources of minerals. Calves (n=43) had body weight and blood collected at birth (prior to suckling), 12 hours post parturition, and 30 days of age. A subset of calves that were evaluated at calving were used to collect serum and liver samples at 115±1.5 days of age and at weaning (205±1.5 days of age). Blood was collected from calves via jugular venipuncture and processed for serum collection. Serum trace mineral analysis for cobalt (Co), copper (Cu), iron (Fe), manganese (Mn), molybdenum (Mo), selenium (Se), and zinc (Zn) was carried out using ICP-Spectroscopy-MS (DCPAH, Lansing, MI). All calves (n=191) were used to calculate weaning average daily gain. Calf mineral status data for 0 to 24 hour post-partum was analyzed using the Mixed model of SAS. Repeated measures analysis was conducted using mineral source treatment, breed, time, and their interactions were included in the model. Means for the 0 to 24 hour period are presented. Mineral status data for day 30 were analyzed using the Mixed model with treatment and breed as fixed effects in the model. Day 115 and weaning data were analyzed using repeated measures in the Mixed procedure of SAS. The fixed effects of mineral source treatment, breed, day, and appropriate interactions were used. Some minerals data required log transformation for analysis. Pearson correlations were determined for

serum and liver trace mineral concentrations. Significance was determined at P 0.05 and a trend 0.05 0.10. Results Time affected (P<0.02) calf serum trace mineral status during the 0 to 24 hour sampling period; with the exception of Co which was not affected by time and Mn which was undetectable. Table 2 presents the 0 to 24 hour mean calf serum trace mineral status. During the 0 to 24 hour period there were no treatment x breed interactions for any of the trace minerals analyzed. Selenium concentration in either whole blood or serum was greater (P<0.001) for ORG compared to ING calves. Concentrations of Co and Mo were not affected (P 0.12) by treatment or breed. In contrast, Cu and Zn concentrations were greater (P<0.05) in than, whereas calves tended to have greater (P=0.09) Fe concentration than calves after birth. The mean mineral status across all calves indicated calves were deficient in Mn and Cu, but marginal to adequate for the remaining trace minerals. Similar to the 0 to 24 hour sampling period, day 30 Se concentration (Table 3) in whole blood and serum was greater (P<0.001) for ORG compared ING calves. Additionally, serum Se concentration was greater (P=0.05) in compared calves. Calves from dams supplemented with ING had greater (P=0.02) Co concentration than ORG calves. Concentrations of Cu, Fe, Mn, Mo, and Zn on day 30 did not differ between treatments (P 0.37) or breed (P 0.42). On day 30 all trace mineral were indicated to be marginal to adequate, Fe concentrations were determined to be high in status across all treatments. Serum Co and Zn were affected (P<0.01) for day 115 and weaning, Co concentrations increased from day 115 to weaning whereas Zn concentrations decreased with time. Organic calves had greater (P<0.001) serum Se concentrations (29.8 ng/ml) compared to ING calves (18.53 ng/ml), while calves had greater (P<0.001, 28.90 ng/ml) Se concentration compared to calves (19.99 ng/ml). Calf Co and Mo concentrations demonstrated breed differences, primarily driven by greater concentrations at weaning for Co for and Mo for. Liver concentrations of trace minerals (Table 5) were affected (P 0.01) day for all minerals. Liver concentrations increased for Co, Cu, Mn, Mo, Se, and Zn; but decreased for Fe. Angus calves had greater (P 0.05) Co, Fe, and Zn liver concentrations than, whereas calves had greater (P<0.05) Mn concentration than calves. Mineral source treatment increased (P 0.01) Mn and Se concentrations in ORG calves compared to ING calves. Mineral source did not affect the liver concentration of Co, Cu, Mo, or Zn in calves at day 115 or weaning. Correlations between serum and liver concentrations were generally poor except for Co and Se (Co, r 2 =0.88; Cu, r 2 =0.00; Fe, r 2 =0.12; Mn, r 2 =0.08; Mo, r 2 =0.04; Se, r 2 =0.58; Zn, r 2 =-0.12). Serum and liver concentrations are both valid means to assess trace mineral status, however for most trace minerals there are poor correlations between measures implying that one or the other assessment should be used, but comparisons between means are not valid. Literature Cited AAFCO. 2000. Official Publication. Association of American Feed Control Officials, Inc. Du, M., et al. 2010. J. Anim. Sci. 88:E51-E60. Larson, D. M., et al. 2009. J. Anim. Sci. 87:1147-1155. Underwood, K. R., et al. 2010. 86:588-593. Zhu, M. J, et al. 2006. J. Pysiol. 575:241-250.

Table 1. Composition of trace mineral supplement provided to cows Component Inorganic Organic Dry matter, % 88.5 88.5 Crude protein, % 14.9 14.9 Total digestible nutrients, % 70.6 70.3 Ca, % 0.21 0.19 P, % 1.00 0.99 Mg,% 0.46 0.46 K, % 1.36 1.35 S, % 0.22 0.16 Co, ppm 19.1 16.7 Cu, ppm 260 150 I, ppm 53.8 20.1 Fe, ppm 407 406 Mn, ppm 367 284 Se, ppm 7.1 6.8 Zn, ppm 892 526 Vitamin A, IU/lb 31,606 31,606 Vitamin D 3, IU/lb 2,835 2,835 Vitamin E, IU/lb 301 301 Table 2. Effect of maternal trace mineral supplement source on 0 to 24 hour post-partum calf trace mineral serum and whole blood (WB) concentrations SEM Trt 2 Mean Status 3 Co, μg/ml 0.32 0.40 0.41 0.43 0.04 0.18 0.27 0.44 M-A Cu, μg/ml 0.23 0.27 0.23 0.27 0.01 0.74 0.003 0.74 D Iron, μg/ml 93.67 67.48 80.58 72.64 8.16 0.63 0.05 0.28 A Mn, μg/ml -- -- -- -- -- -- -- -- -- Mo, ng/ml 6.27 5.13 7.47 6.05 0.80 0.20 0.12 0.87 A Se-WB, ng/ml 116.30 121.12 154.31 144.48 4.59 <0.001 0.59 0.12 A Se, ng/ml 47.27 45.64 62.12 52.82 2.39 <0.001 0.03 0.12 A Zn, μg/ml 0.58 0.74 0.53 0.65 0.08 0.38 0.09 0.79 M 1 ING = inorganic, ORG = organic, = Angus, = Brangus. 3 Trace mineral status, A = adequate, M = marginal, D = deficient. Reference ranges for TM status from Herdt and Hoff, 2011. Vet. Clin. Food Anim. 27: 255 283.

Table 3. Effect of maternal trace mineral supplement source on 30-day post-partum calf trace mineral serum and whole blood (WB) concentrations SEM Trt 2 Mean Status 3 Co, μg/ml 1.56 1.60 0.46 0.65 0.45 0.02 0.80 0.87 A Cu, μg/ml 0.62 0.39 0.73 0.94 0.57 0.40 0.98 0.58 A Fe, μg/ml 215.2 241.6 198.3 195.4 36.1 0.37 0.74 0.68 H Mn, μg/ml 1.77 1.87 2.32 1.61 0.39 0.70 0.42 0.28 A Mo, μg/ml 5.11 5.05 5.59 4.87 0.88 0.86 0.65 0.70 M-A Se-WB, ng/ml 96.30 94.55 133.82 125.91 4.73 <0.001 0.30 0.51 M Se, ng/ml 34.5 32.27 46.91 42.45 1.69 <0.001 0.05 0.50 M-A Zn, μg/ml 1.10 1.25 1.12 0.47 0.60 0.37 0.55 0.34 M 1 ING = inorganic, ORG = organic, = Angus, = Brangus. 3 Trace mineral status, A = adequate, M = marginal, D = deficient. Reference ranges for TM status from Herdt and Hoff, 2011. Vet. Clin. Food Anim. 27: 255 283. Table 4. Effect of maternal trace mineral supplement source on 115-day post-partum and weaning calf serum trace mineral concentrations SEM Trt 2 Day x Day Co, ng/ml d 115 0.53 0.39 0.50 0.23 0.17 0.11 0.03 <0.01 0.59 weaning 2.32 0.71 0.79 0.77 0.73 Cu, ng/ml d 115 0.58 0.57 0.61 0.61 0.04 0.48 0.68 0.91 0.08 weaning 0.61 0.56 0.54 0.65 0.04 Fe, ng/ml d 115 190.3 100.5 154.9 166.3 13.36 0.24 0.38 0.27 0.17 weaning 135.7 131.9 122.8 163.4 10.05 Mn, ng/ml d 115 2.25 1.40 1.49 2.90 0.32 0.24 0.67 0.13 0.05 weaning 1.73 1.45 1.67 1.73 0.32 Mo, ng/ml d 115 2.02 6.47 3.36 2.27 1.01 0.17 0.005 0.37 <0.001 weaning 2.57 1.77 2.13 9.66 1.01 Se, ng/ml d 115 22.13 17.33 36.87 23.42 1.14 <0.001 <0.001 0.20 0.10 weaning 20.98 13.67 35.62 25.56 0.99 Zn, ng/ml d 115 0.97 0.87 0.92 0.99 0.03 0.34 0.65 <0.001 0.08 weaning 0.79 0.77 0.87 0.83 0.03 1 ING = inorganic, ORG = organic, = Angus, = Brangus.

Table 5. Effect of maternal trace mineral supplement source on 115-day post-partum and weaning calf liver trace mineral concentrations SEM Trt 2 Day x Day Co, μg/g d 115 0.12 0.10 0.12 0.08 0.02 0.13 0.01 0.002 0.15 weaning 0.36 0.14 0.19 0.14 Cu, μg/g d 115 75.23 58.10 84.06 64.76 8.35 0.60 0.29 0.06 0.19 weaning 78.58 96.59 125.29 73.31 10.92 Fe, μg/g d 115 731 581 588 415 37.1 0.13 0.01 0.005 0.29 weaning 543 372 463 402 28.8 Mn, μg/g d 115 6.93 6.15 6.90 10.53 0.42 0.01 0.05 0.01 0.26 weaning 8.98 9.12 9.06 10.63 0.42 Mo, μg/g d 115 1.72 1.88 1.89 1.87 0.12 0.93 0.22 <0.001 0.86 weaning 2.33 2.90 2.34 2.66 0.12 Se, μg/g d 115 0.05 0.78 0.90 0.64 0.05 0.007 0.41 0.03 0.21 weaning 0.40 0.33 0.83 0.60 0.05 Zn, μg/g d 115 125.7 102.3 121.2 115.7 3.59 0.30 0.04 0.03 0.56 weaning 144.0 131.9 122.8 118.6 3.99 1 ING = inorganic, ORG = organic, = Angus, = Brangus. Table 6. Effect of maternal trace mineral supplement source on calf body weight Mineral source breed combination 1 SEM Trt 2 Day x Day Birth weight, lb 3 77 79 79 80 1.9 0.48 0.34 -- 0.77 30-d ADG, lb/d 3 2.14 1.79 0.44 0.68 0.40 0.002 0.89 -- 0.48 Day 115, lb 332 328 326 329 7.36 0.18 0.76 <0.01 <0.01 Weaning, lb 468 484 500 494 7.36 ADG, lb/day 3 1.92 2.07 1.98 2.01 0.04 0.02 0.82 -- 0.13 1 ING = inorganic, ORG = organic, = Angus, = Brangus. 3 Treatment x breed interaction only.