CASE STUDY: Effects of Supplementing Injectable Trace Minerals on Dairy Calf Performance

The Professional Animal Scientist 26 (2010):667–671

©2010 American Registry of Professional Animal Scientists

CSupplementing S : Effects of Injectable ASE

TUDY

Trace Minerals on Dairy Calf Performance T. D. Nennich,*1,2 PAS, J. R. Crenwelge,† B. D. Lambert,*† PAS, N. M. Cherry,* and E. R. Jordan,‡ PAS *Texas AgriLife Research and Extension Center at Stephenville, Texas A&M University System, Stephenville 76401; †Department of Animal Sciences, Tarleton State University, Stephenville, TX 76401; and ‡Texas AgriLife Extension Service, Texas A&M University System, Dallas 75252

ABSTRACT Calves from a commercial dairy operation were studied to determine if an alternative method of supplying trace minerals would improve calf growth and mineral status. The objective of these trials was to determine the effects of supplementing trace minerals with an injectable mineral product (MIN), containing 16 mg/mL Cu, 10 mg/mL Mn, 5 mg/mL Se, and 48 mg/mL Zn, on ADG and blood and liver mineral concentrations of preweaned dairy calves. In trial 1, 123 Holstein heifers were randomly assigned to either the control or the MIN treatment. Calves on the MIN treatment (n = 60) received a 1-mL injection of MIN at 1 d of age. Calves were weighed on d 3 ± 2 and again on d 42 ± 3 to determine ADG. Trial 2 included 10 Holstein bull calves, with MIN calves (n = 5) receiving a 1-mL injection of MIN at d 4. Blood and liver samples were collected on d 3 and 43. In trial 1, Corresponding author: tnennich@purdue. edu 2 Present address: Department of Animal Sciences, Purdue University, West Lafayette, IN 47907. 1

ADG (0.29 and 0.30 kg/d for control and MIN, respectively) and weaning weights (49.0 and 50.1 kg for control and MIN, respectively) were similar (P > 0.10). In trial 2, ADG for control and MIN calves were similar (P > 0.10) and averaged 0.49 and 0.50 kg/d, respectively. Blood and liver mineral concentrations were similar (P > 0.10) for control and MIN calves. In these trials, an injection of this mineral product at birth did not significantly affect ADG or blood and liver mineral concentrations of dairy calves. Key words: calf, dairy, trace mineral

INTRODUCTION Concentrations of trace minerals in the blood and liver of dairy calves at birth are dependent on nutritional status of the dam (Hidiroglou and Knipfel, 1981; Abdelrahman and Kincaid, 1993). The most limiting trace elements for a fetus are Cu, Mn, Zn, and Se, and a deficiency of one of these elements can reduce the tissue reserves of these trace minerals in neonatal calves (Abdelrahman and Kincaid, 1993). Trace mineral

needs of milk-fed dairy calves are assumed to be met through intake of milk or milk replacer and calf starter. Trace minerals participate in a wide range of body functions; most of them as components of important enzymatic systems that involve immune, metabolic, and reproductive functions (Graham, 1991; Ferguson, 1996; NRC, 2001), thus marginal levels of trace minerals have the potential to negatively affect animal health. Alternate methods of supplementing trace minerals may provide an opportunity to improve animal growth and performance. Proper mineral nutrition and supplementation is essential to animal health and growth in young calves. One of the major disadvantages of the use of dietary mineral supplements is that they may not be absorbed properly due to interactions with other nutrients in the digestive tract (King, 1971; Stake, 1977; Suttle, 1986). High dietary intakes of other minerals including Zn, Fe, Mo, and S have affected Cu utilization in adult cattle (McDowell, 1992). High levels of dietary Zn can result in reduced concentrations of Cu in plasma and

668 liver of cattle and sheep (Ott et al., 1966; Kincaid et al., 1976; Kellogg et al., 1989). Furthermore, excess intake of dietary Mo can inhibit uptake and utilization of Cu through the formation of thiomolybdates in the rumen and was found to mobilize Cu from the liver in ewes (Kincaid and White, 1988). Due to concerns with mineral absorption and interactions, the use of injectable trace minerals has become an area of interest. The objective of these trials was to evaluate the effect of supplementing trace minerals using an injectable mineral product (MIN) on dairy calf performance. Two separate trials were conducted to evaluate the growth rate of young calves during the first 6 wk of life and to determine if a single injection of a mineral supplement affected blood or liver mineral concentrations.

MATERIALS AND METHODS All protocols were approved by the Tarleton State University Animal Care and Use Committee. Calves used for both trials were obtained from a commercial dairy operation in Gustine, Texas. In both trials, calves were housed in 1.2 by 2.4 m fiberglass calf hutches with an exterior panel (1.2 by 1.8 m) to allow room for mobility.

Trial 1 In the first trial, 123 Holstein heifers were assigned to treatment according to ear tag number, with oddnumbered calves assigned to control and even-numbered calves allotted to the treatment. Heifer calves were born at a commercial dairy operation and transported to a calf and heifer raising operation, located 12.6 km away, within 1 d of birth. The trial took place from early October 2006 to late November 2006 and was a completely randomized design. Treatment calves (n = 60) received a subcutaneous 1-mL injection of MIN (Mineral Max II 3 to 1 Formula, Walco International, Westlake, TX), containing 16 mg/mL Cu, 10 mg/mL Mn, 5 mg/mL Se, and 48 mg/mL Zn, at 1 d of age

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Table 1. Analysis of milk replacer and calf starter fed to 123 Holstein heifer calves Item DM, % CP, % of DM NDF, % of DM ADF, % of DM Ash, % of DM Ca, % of DM P, % of DM Fe, mg/kg Mn, mg/kg Zn, mg/kg Cu, mg/kg

Calf starter

Milk replacer

85.7 20.3 21.2

85.7 25.2 0.3

10.1

0.1

9.9

14.2

1.47 0.71 282 108 109 46

1.06 1.01 94 28 44 26

and were weighed at 3 ± 2 and 42 ± 3 d of age to determine ADG. Initial weights were obtained using a tripod, sling, and spring scale, whereas final weights were measured with a portable, digital scale. All weights were deemed accurate to ±0.5 kg. The calves in this trial were fed using 4-L galvanized pails located adjacent to each other. Calves were fed approximately 0.454 kg/d of a 22% CP, 20% fat milk replacer (Purina Mills, St. Louis, MO) in liquid volume of 4 L and had ad libitum access to a texturized calf starter (18% CP, as fed; Purina Mills) and clean drinking water at all times during the experiment. Feed and milk replacer samples were collected on a weekly basis, frozen, and composited every 2 wk before analysis. Feed and milk starter samples were analyzed using wet chemistry methods by Cumberland Valley Analytical Services Inc. in Hagerstown, Maryland. Nutrient composition of the milk replacer and calf starter are provided in Table 1.

Trial 2 Calves in trial 2 were sourced from the same commercial dairy operation as those in trial 1 and were housed at the Tarleton State University Dairy Center in Stephenville, Texas, after

being transported 63.7 km. Ten intact Holstein male calves were assigned to 1 of 2 treatments in a completely randomized design. Calves assigned to the MIN treatment (n = 5) received a 1-mL injection of MIN at 4 d of age. Calves were initially weighed at 4 d of age and again at 43 d of age at the completion of the trial. This experiment was conducted from mid-December 2006 to late January 2007. A portable, digital scale with an accuracy of ±0.5 kg was used to determine BW of calves during this experiment. Calves were bottle fed approximately 4 L of nonsaleable milk per day from the Tarleton State University Dairy and had ad libitum access to calf starter (Table 1). Calves were supplied clean drinking water in black plastic 4-L pails that were located adjacent to the starter pail at all times during the experiment. Pretreatment blood samples and liver biopsies were collected from all calves 1 d before treatment administration, on d 3 and again at the completion of the trial on d 43. Liver biopsies were obtained as described by Chapman et al. (1963). An area in the caudo-thoracic region was clipped and sanitized using an iodophor scrub. Following administration of local anesthesia at the biopsy site, a small incision was made to allow the biopsy trocar to be inserted through the body wall and peritoneum between the 12th and 13th ribs. The trocar was directed into the liver parenchyma, where a liver sample (~3 g) was collected. Following the biopsy, the site of incision was sutured and an antiseptic agent was applied. In addition, an antibiotic injection was given for 3 d following the procedure to prevent infection. Approximately 6 mL of blood was collected via the jugular vein and divided between two 5-mL blood tubes, one containing heparin and one without. Tubes that did not contain heparin were centrifuged, and serum was removed and placed in individual containers for Cu, Zn, and Mn analysis. Whole blood samples were collected for analysis of glutathione peroxidase (GSH-Px). Liver, serum,

Injectable trace mineral supplementation for dairy calves

Table 2. Birth weights, weaning weights, ADG, age at weaning, and death rates for 123 Holstein heifer calves receiving either no treatment (control) or 1 mL of a mineral supplement (MIN) containing 16 mg/mL Cu, 10 mg/mL Mn, 5 mg/mL Se, and 48 mg/mL Zn at 1 d of age Control

MIN

Item

n

Mean (SE)

n

Mean (SE)

P-value

Birth weight, kg Weaning weight, kg ADG, kg Age at weaning, d Death rate, %

63 56 56 56 7

36.2 (0.60) 49.0 (0.87) 0.30 (0.016) 42.2 11.1

60 55 55 55 5

36.9 (0.61) 50.1 (0.88) 0.31 (0.016) 42.0 8.3

0.41 0.37 0.67 — —

and whole blood samples were placed on ice and sent to Colorado Veterinary Diagnostic Laboratories in Fort Collins, Colorado, for analyses. Copper, zinc, and liver manganese were analyzed using flame atomic absorption spectrometry (Varian Inc., Palo Alto, CA). Serum manganese levels were estimated using graphite furnace atomic absorption spectrometry (Varian Inc.). Selenium was analyzed via hydride vapor generation atomic absorption spectrometry (Varian Inc.), and GSH-Px was determined through an enzyme rate reaction with spectrophotometric (Varian Inc.) determination of GSH-Px activity. Data from each trial were analyzed separately as a completely randomized design. In both trials, the data were analyzed using the MIXED procedure of SAS (SAS Institute, 2003) with calf as the experimental unit. The model contained the effects of treatment. In trial 2, blood and liver mineral concentrations from the original collection on d 4 were used as a covariate.

RESULTS AND DISCUSSION Supplementation of trace minerals to neonatal dairy calves with an injectable product did not affect growth performance of calves in these trials. In trial 1, the ADG and weaning weights of calves receiving the mineral injection were similar to those of control calves (Table 2); however, the ADG of calves (0.30 kg/d) in trial 1 were much lower than expected.

Average daily gains for control and MIN calves in trial 2 were similar (P > 0.10), averaging 0.49 and 0.50 kg/d, respectively. Weaning weights of calves in trial 2 averaged 66.8 kg and were similar between treatments (P > 0.10). Weight gains of calves were closer to expected levels in trial 2, with the ADG of the calves in trial 2 exceeding those from trial 1. Because calves were from the same farm and colostrum management was similar for calves in both trials, the lower rates of gain in trial 1 may have been due to maternal heat stress, lower calf birth weights, and potential fetal immune suppression. Previous studies also reported similar growth rates for calves supplemented with various levels of trace minerals as compared with control calves (Kincaid et al., 1986; Kincaid et al., 1997). Feeding different concentrations and sources, either organic or inorganic sources, of Zn supplements to 6-wk-old calves did not affect ADG (Kincaid et al., 1997). Similarly, Kincaid et al. (1986) found that feeding older calves (>12 wk of age) organic Cu sources increased plasma Cu concentration after 12 wk of supplementation but did not result in any improvements in calf growth rates. Furthermore, death rates in trial 1 were 8 and 11% for the MIN and control calves, respectively (Table 2). By supplementing these calves, there was hope of eliminating or reducing death losses, but there were not enough calves in the trial to evaluate

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the effects of the MIN supplement on death rates. Calves in these trials were on standard milk feeding programs and received 4 L of milk or milk replacer per day. Due to the rearing program, energy and protein may have been the first limiting factors for ADG in these calves, thus the quantity of micronutrients may not have been the limiting factor in the diets of these calves. The results of this study would apply only to traditional rearing programs and may be different for accelerated calf feeding programs. There were no significant treatment differences observed in trial 2 between blood and liver mineral concentrations at d 43 for control calves or dairy calves receiving the MIN supplement (Table 3). A variety of factors including minerals in milk, calf starter mineral levels, and stress can affect mineral concentrations in the blood and liver of calves. At the start of the trial, blood and liver levels of Cu, Zn, Se, and GSH-Px were in the normal range for calves, although liver Mn levels were lower than expected. Because mineral levels in calves were adequate at birth, additional supplementation may not have elicited a biological response. Also, the minerals present in the MIN supplement either were not adequately absorbed by the calves or were not supplemented in great enough quantities to result in an increased long-term storage of the minerals over the milk feeding period of the calves. In trial 2, liver Cu concentrations of the calves at 43 d of age (Table 3) were greater than values reported by Alfaro et al. (1988) and Kincaid et al. (1997) and less than those reported by Neathery et al. (1991) for 12-wkold calves. The liver Cu concentrations were expected to be high, because the liver of newborn ruminants normally contains high (>200 mg Cu/kg liver DM) concentrations of Cu (Hidiroglou and Williams, 1982; Branum et al., 1998). The Cu status of calves is affected by maternal Cu status (Bremner and Dalgarno, 1973; Gooneratne et al., 1989). Kincaid et al. (1986) reported that calves fed

670 an organic Cu supplement did have greater concentrations of Cu in liver after being supplemented 12 wk as compared with control or an inorganic copper supplement. The level of Cu supplementation in this study was low compared with typical liver Cu concentrations, and a single dose of supplemented Cu did not appear to be adequate to change the storage of Cu in the liver in calves in this study. The concentrations of plasma (Table 3) Zn in this trial were similar to values reflected in the literature; previous research shows the levels of Zn are relatively high in newborn calves but drop to 1.2 μg/mL by 12 wk of age (Kincaid and Hodgson, 1989). Kincaid et al. (1997) reported an increase in serum Zn for calves fed a starter containing 300 mg Zn/

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kg DM of an organic Zn supplement when compared with an inorganic Zn supplement included in the starter at a similar supplementation rate, but no differences in serum Zn concentrations were seen when organic Zn supplements were included in the diet at a rate of 150 mg Zn/kg DM. Concentrations of Zn in the liver of calves in trial 2 ranged from 70 to 82.8 mg/ kg for the control and MIN groups, respectively, which were lower than previously reported values of 100 to 400 mg/kg DM (Puls, 1994), 136 mg/ kg DM of liver Zn for calves dosed orally with Zn (Neathery et al., 1991), and 208 to 390 mg/kg DM reported by Kincaid et al. (1997). When comparing liver Mn concentrations of calves in trial 2 to previously reported values (Ho et al., 1984;

Table 3. Blood, serum, and liver mineral concentrations of Holstein bull calves at the start of the trial and at 43 d of age1 Item Initial measurements   Whole blood    Glutathione peroxidase, mmol/L per second   Serum    Cu, mg/kg    Mn, mg/kg    Zn, mg/kg   Liver    Cu, mg/kg    Mn, mg/kg    Zn, mg/kg    Se, mg/kg Measurements of calves at 43 d of age   Whole blood    Glutathione peroxidase, mmol/L per second   Serum    Cu, mg/kg    Mn, mg/kg    Zn, mg/kg   Liver    Cu, mg/kg    Mn, mg/kg    Zn, mg/kg    Se, mg/kg

Control    

416

MIN    

400

SE    

39.2

P-value    

0.79

  0.36 0.001 2.55   261 3.14 375.8 2.58     483

    0.45 0.040 0.001 0.0001 1.99 0.140     292 35.7 4.49 0.272 269.7 65.30 2.69 0.362         531 32.7

  0.15 0.35 0.02   0.55 0.01 0.28 0.83     0.37

  0.40 0.001 0.99   408.9 6.47 70.03 3.83

    0.42 0.02 0.001 0.0002 1.02 0.03     545.8 90.9 7.43 0.83 82.77 7.12 5.23 1.20

  0.50 0.41 0.59   0.37 0.53 0.30 0.47

Treatment calves (MIN) were injected with 1 mL of a mineral supplement, containing 16 mg/mL Cu, 10 mg/mL Mn, 5 mg/mL Se, and 48 mg/mL Zn, at 4 d of age. Measurements of calves at 43 d of age were adjusted using initial measurements as a covariate.

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Jenkins and Hidiroglou, 1991), liver Mn concentrations of 3.14 and 4.49 mg/kg of calves at the start of the trial in the control and MIN groups, respectively, are lower than the 10 to 24 mg/kg previously reported for calves with adequate concentrations of Mn in the liver (Puls, 1994). The liver Mn values for calves at the end of the trial (Table 3) were similar to liver Mn values of 7.0 to 9.6 mg/kg reported by Neathery et al. (1991) for weaned Holstein bull calves. The NRC (1989) reported that 40 mg Mn/kg DM should be adequate for all classes of cattle with no requirements determined. Selenium concentrations, although tested in the liver, are better determined by GSH-Px concentrations in the blood. Results from calves in trial 2 (Table 3) found GSH-Px levels to average 483 and 531 mmol/L per second for control and MIN calves, respectively. In liver samples collected on d 43 (Table 3), levels of Se in the liver were 3.83 and 5.23 mg/kg for control and MIN groups, respectively. These liver Se concentrations were slightly greater than recommended Se liver concentrations of 2.2 mg/kg DM for normal growth and health in calves (Abdelrahman and Kincaid, 1995). Although the liver Se values of calves in this study were greater than recommendations, the liver Se levels were similar to levels reported by Jenkins and Hidiroglou (1986) in which calves receiving Se supplements of either 3 or 5 mg/kg DM per day had average Se liver concentrations of 4.74 and 9.90 mg/kg, respectively.

IMPLICATIONS Supplementing calves with a 1-mL injection of a trace mineral product, containing 16 mg/mL Cu, 10 mg/mL Mn, 5 mg/mL Se, and 48 mg/mL Zn, at birth did not significantly affect the ADG or blood and liver mineral concentrations of dairy calves at weaning in these studies. Supplementing trace minerals to dairy calves at these levels was either not adequate to increase the mineral status of the calves, or the background mineral

Injectable trace mineral supplementation for dairy calves

concentrations of the calves were adequate to support growth rates in these studies and were not a limiting nutritional factor. Additional studies to determine the effects of supplementing dry cows with an injectable mineral supplement or studies in which calves receive multiple injections of mineral supplements need to be evaluated.

ACKNOWLEDGMENTS We would like to thank Walco International, Westlake, Texas, for financial support of the study.

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Ferguson, J. D. 1996. Diet, production, and reproduction in dairy cows. Anim. Feed Sci. Technol. 59:173.

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