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J. Anim. Sci. 2003. 81:1357-1362
© 2003 American Society of Animal Science

Effect of copper source and level on performance and copper status of cattle consuming molasses-based supplements1,2

J. D. Arthington*,3, F. M. Pate* and J. W. Spears{dagger}

* University of Florida, Institute of Food and Agricultural Sciences, Range Cattle Research and Education Center, Ona 33865 and and {dagger} Department of Animal Science and Interdepartmental Nutrition Program, North Carolina State University, Raleigh 27695

3 Correspondence: 3401 Experiment Station (phone: 863-735-1314; fax: 863-735-1930; E-mail:
jdarthington{at}mail.ifas.ufl.edu).


   Abstract
Top
 Abstract
Introduction
 Materials and Methods
 Results and Discussion
 Implications
 Literature Cited
 
Two studies were conducted to evaluate the availability of dietary Cu offered to growing beef cattle consuming molasses-based supplements. In Exp. 1, 24 Braford heifers were assigned randomly to bahiagrass (Paspalum notatum) pastures (two heifers/pasture). Heifers were provided 1.5 kg of TDN and 0.3 kg of supplemental CP/heifer daily using a molasses–cottonseed meal slurry. Three treatments were randomly assigned to pastures (four pastures/treatment), providing 100 mg of supplemental Cu daily in the form of either CuSO4 (inorganic Cu) or organic-Cu. A third treatment offered no supplemental Cu (negative control). Heifer BW was collected at the start and end of the study. Jugular blood and liver samples were collected on d 0, 29, 56, and 84. In Exp. 2, 24 Brahman-crossbred steers were fed the same molasses–cottonseed meal supplement at the same rates used in Exp. 1. Steers were housed in individual pens (15 m2) with free-choice access to stargrass (Cynodon spp.) hay. Four Cu treatments were assigned to individual steers (six pens/treatment) providing 1) 10 ppm of Cu from an organic source; 2) 10 ppm Cu from Tri-basic Cu chloride (TBCC); 3) 30 ppm of Cu from TBCC; or 4) 30 ppm of Cu, a 50:50 ratio of TBCC and organic Cu. Body weights and jugular blood and liver samples were collected on d 0, 24, 48, and 72. In Exp. 1, liver Cu concentrations did not differ between heifers supplemented with inorganic and organic Cu. Each source resulted in increased (P < 0.05) liver Cu concentrations compared with the unsupplemented control. Plasma ceruloplasmin concentrations were higher (P < 0.05) for Cu-supplemented heifers, independent of Cu source. Heifer ADG tended (P = 0.11) to increase with Cu supplementation compared with the unsupplemented control. In Exp. 2, liver Cu was greater (P < 0.05) on d 24, 48, and 72 for steers consuming 30 vs. 10 ppm of Cu. Steers supplemented with organic Cu had lower DMI than steers supplemented with 10 or 30 ppm of TBCC. These data suggest that the inorganic and organic Cu sources evaluated in these studies were of similar availability when offered in molasses supplements. A dietary Cu concentration greater than 10 ppm might be necessary to ensure absorption in beef cattle fed molasses-based supplements.

Key Words: Cattle • Copper • Molasses


   Introduction
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 Abstract
 Introduction
Materials and Methods
 Results and Discussion
 Implications
 Literature Cited
 
In a previous series of studies (Arthington and Pate, 2002), heifers provided molasses-based supplements had lower liver Cu concentrations compared with heifers consuming a similar amount of Cu provided in a corn-based supplement. High S concentrations, naturally found in molasses, were suggested to be the most likely reason for this reduction in Cu availability. A second study within this series supported this supposition, in that heifers consuming corn-based supplements fortified with S had lower liver Cu concentrations compared with heifers consuming corn-based supplements without additional S. The supplemental S level used in that experiment was equal to the amount consumed by heifers provided molasses-based supplements. This S effect is possibly the result of decreased Cu absorption due to the formation of ruminal thiomolybdates (Mason, 1990). The formation of thiomolybdates is directly dependent on available dietary S, and S intake is a major factor influencing the sensitivity of ruminants to Mo (Mason, 1981). Sulfur may also decrease Cu availability independent of Mo. Suttle (1974) found that the addition of both organic and inorganic S to the diets of Cu-deficient sheep decreased the rate and extent of Cu repletion. These responses were attributed to the formation of insoluble Cu sulfide complexes in the gut. These data suggested that an increase of dietary S from 0.1 to 0.4% of the total diet may result in a 50% increase in the overall dietary Cu requirement.

Organic Cu may be more bioavailable than CuSO4 when beef cattle consume high-S forages (Arthington et al., 2002). In other studies, an inorganic source of Cu, tri-basic Cu chloride (TBCC; Micronutrients Inc., Indianapolis, IN), was suggested to be more bioavailable compared with CuSO4 (J. W. Spears, unpublished data). Therefore, the objective of the current studies was to investigate the effect of Cu level and source on Cu status of growing beef cattle fed molasses-based supplements.


   Materials and Methods
Top
 Abstract
 Introduction
 Materials and Methods
Results and Discussion
 Implications
 Literature Cited
 
Animal Care, Handling, and Diet

The animals utilized in these experiments were cared for by acceptable practices (FASS, 1999) and the protocol was approved by the University of Florida, Institutional Animal Care and Use Committee (#A759). Liver biopsy collections were performed by a trained technician using techniques previously described (Arthington and Corah, 1995).

In Exp. 1, 24 nonpregnant Braford heifers (11 to 14 mo of age; average BW = 353 ± 13.1 kg) were randomly assigned to bahiagrass (Paspalum notatum) pastures of equal size (1.22 ha; two heifers/pasture). Initial liver Cu concentrations were considered normal for Florida beef cattle (normal liver Cu concentration = 100 to 300 ppm on a DM basis; Ammerman, 1969). Heifers were provided a molasses supplement fortified with cottonseed meal (20%, as-fed basis) formulated to provide, on average, 1.5 kg of TDN and 0.3 kg of CP/heifer daily. Supplements were fed three times weekly (3.5 and 0.7 kg of TDN and CP/heifer, provided on Monday, Wednesday, and Friday). Three Cu treatments were randomly assigned to pastures (four pastures/treatment). Treatments consisted of 1) control (no supplemental Cu), 2) inorganic Cu (CuSO4), or 3) organic Cu (Availa-Cu, Zinpro Corporation, Eden Prairie, MN). Each supplemental Cu source was provided at a rate of 100 mg/d, corresponding to approximately 10 ppm assuming a DMI of 2.5% of BW. Treatments were mixed directly into the molasses supplements in conjunction with a complete mineral mixture (Table 1Go). Complete consumption of the supplement was achieved within 24 to 36 h. To assess the effect of supplement composition on animal performance, individual heifer weights were collected at the start and conclusion of the study, following a 12-h shrink. To assess the effect of Cu source on liver Cu concentrations and plasma ceruloplasmin activity, liver biopsies and jugular blood collections were performed on d 0, 29, 56, and 84.


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  		Table 1. Mineral composition of molasses slurry and forage (DM basis) from Experiments 1 and 2a
 
In Exp. 2, 24 Brahman x Angus crossbred steers (average BW = 249 ± 16.2 kg) were randomly allocated into individual covered pens (15 m 2; one steer/pen). All steers received a common molasses–cottonseed meal supplement (20% slurry) formulated to provide, on average, 1.5 kg of TDN and 0.3 kg of CP/steer daily. Supplements were fed three times weekly (3.5 and 0.7 kg of TDN and CP/steer, provided on Monday, Wednesday, and Friday). Complete consumption of the supplement offered was achieved within 24 to 36 h. Four Cu treatments were randomly assigned to individual pens (six pens/treatment). Copper treatments were formulated using an estimated DMI of 2.25% BW for supplemental dietary Cu concentrations of 10 ppm (56 mg of Cu/d) and 30 ppm (168 mg of Cu/d). Individual treatments were 1) 10 ppm of Cu from organic Cu (Availa-Cu); 2) 10 ppm of Cu from TBCC (Micronutrients Inc.), 3) 30 ppm of Cu from TBCC, or 4) 30 ppm of Cu (50:50 ratio of TBCC and organic Cu). Treatments were mixed directly into the molasses supplements in conjunction with a complete mineral mixture (Table 1Go). To assess the effect of Cu level and source on animal performance, liver Cu concentration, and plasma ceruloplasmin activity, unshrunk BW, jugular blood, and liver biopsy collections were on d 0, 24, 48, and 72. All steers were offered free-choice access to ground stargrass hay (Cynodon spp.) To determine the effect of dietary treatment on DMI, daily forage offered and refusal was determined. Mean DMI was calculated on 6-d intervals for every steer throughout the 72-d experiment.

Feed, Plasma, and Liver Analysis

Random samples of bahiagrass (hand-clipped), hay, molasses, and cottonseed meal were collected and analyzed for mineral concentration by a commercial laboratory (SDK laboratories, Hutchinson, KS; Table 1Go) using inductively coupled plasma-atomic emission spectroscopy. Following collection of liver biopsy samples, samples were frozen and sent to Michigan State University (Animal Health Diagnostic Laboratory, Lansing, MI) for analysis of trace mineral concentration using inductively coupled plasma-atomic emission spectroscopy as described by Braselton et al. (1997). Blood was collected by jugular venapuncture into heparin-coated, evacuated tubes. Plasma was harvested from blood following centrifugation at 2,400 x g for 20 min and then frozen at -20°C until analyzed for ceruloplasmin concentration using colorimetric procedures (Demetriou et al., 1974).

Statistical Analysis

Statistical analyses of liver mineral and plasma ceruloplasmin concentrations were achieved by ANOVA for a repeated measures experiment within a completely randomized design using the PROC MIXED procedure of SAS (SAS Inst., Inc., Cary, NC). The model statement contained the effects of treatment and day and the interaction for treatment x day. Data were analyzed using the pasture (or pen; Exp. 2) x treatment interaction as random effects. Initial liver Cu concentrations did not differ significantly in either experiment; therefore, these values were used as covariates in the model for analysis of subsequent sampling dates. Statistical analysis for overall change in liver Cu concentration and ADG was achieved by ANOVA for a completely randomized design using the PROC GLM procedure of SAS, with pasture x treatment as the error term. Pasture (or pen; Exp. 2) was the experimental unit. The model statement contained the effect of treatment. When treatment was significant, differences among treatments were compared using single-df orthogonal contrasts. When the treatment x day interaction was significant, data were analyzed by day. Comparisons for Exp. 1 were no Cu vs. Cu and organic Cu vs. inorganic Cu. Comparisons for Exp. 2 were 10 ppm of Cu vs. 30 ppm of Cu, TBCC at 10 and 30 ppm, and treatments with organic Cu vs. treatments without.


   Results and Discussion
Top
 Abstract
 Introduction
 Materials and Methods
 Results and Discussion
Implications
 Literature Cited
 
In Exp. 1, liver Cu concentration was affected by a time x treatment interaction (P < 0.06; Figure 1Go). Initial liver Cu concentrations were similar across treatments; however, on d 29, 56, and 84, heifers supplemented with Cu had greater (P < 0.01) liver Cu concentrations than unsupplemented controls. Liver Cu concentration was similar in heifers supplemented with inorganic and organic Cu sources. Plasma ceruloplasmin concentration was increased with Cu supplementation independent of Cu source (Table 2Go; time x treatment interaction, P = 0.20). Irrespective of Cu source, heifer ADG tended (P = 0.11) to be increased with Cu supplementation compared with control heifers receiving no Cu supplement.



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  		Figure 1. The effect of organic vs inorganic Cu on liver Cu concentration in Exp. 1. Pooled SEM = 26.4 ppm; six pastures/treatment. Values are provided on a DM basis. Treatment x day interaction (P < 0.06); * = P < 0.05.

 

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  		Table 2. Effect of Cu source on heifer average daily gain and plasma ceruloplasmin concentrations, Exp. 1
 
In Exp. 2, liver Cu concentration was affected by a time x treatment interaction (P < 0.01; Table 3Go). Initial liver Cu concentrations were similar across treatments; however, steers consuming Cu at 30 ppm had a greater liver Cu concentration than steers consuming 10 ppm of Cu on d 24, 48, and 72. The overall percentage change in liver Cu was greater (P < 0.01) for steers consuming 30 vs. 10 ppm of Cu (68.1 vs. 11.6%, respectively; SEM = 18.6). Steers consuming TBCC at 30 ppm had a greater (P < 0.05) liver Cu concentration than steers consuming TBCC at 10 ppm on d 48 and 72.


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  		Table 3. Effect of Cu source and level on liver Cu concentrations (DM basis), Exp. 2
 
In Exp. 2, hay DMI increased (P < 0.05) with time for all treatments (Figure 2Go). Steers supplemented with organic Cu had a lesser (P < 0.02) DMI of hay compared with steers supplemented with 10 or 30 ppm of TBCC (Figure 2Go) over the final 36 d (6 intervals) of the study. Despite the difference in DMI, steers consuming organic Cu had a similar (P = 0.26) ADG compared to steers consuming TBCC alone (0.32 and 0.42 kg/d, respectively; SEM = 0.04). Neither a positive nor a negative control was included in Exp. 2; therefore, we were not able to determine if organic Cu was depressing DMI or if TBCC was increasing DMI.



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  		Figure 2. The effect of molasses supplements containing organic Cu (10 ppm of Cu from organic Cu and 30 ppm of Cu from a 50:50 ratio of organic Cu and Tri-basic Cu Chloride [TBCC]) vs. molasses supplements formulated without organic Cu (10 and 30 ppm of Cu from TBCC) on hay DMI; single-df contrast for Cu source (P < 0.02), Exp. 2. Pooled SEM = 0.15 kg/d; 12 animals/treatment. Values are on a DM basis. * = Means differ (P < 0.05).

 
Plasma ceruloplasmin concentrations did not differ among treatments or sampling days in Exp. 2 (average ceruloplasmin concentration over all times = 9.9, 12.5, 12.9, and 12.0 mg/dL for TBCC at 10 ppm, organic Cu at 10 ppm, TBCC at 30 ppm, and TBCC + organic Cu at 30 ppm, respectively; SEM = 0.78). The lack of a negative control, which received no supplemental Cu, made treatment differences in plasma ceruloplasmin concentration unlikely. These data suggest that both Cu sources were of similar availability when offered to cattle in molasses-based supplements.

Trace minerals complexed with organic molecules have been implied to be more bioavailable than inorganic trace minerals (Brown and Zeringue, 1994). Some researchers (Nockels et al., 1993; Rabiansky et al., 1999) have indicated that Cu-lysine may be a more available source of Cu than CuSO4 in cattle. The physiological advantage afforded by organic Cu compounds may be due to the unique coordination chemistry of Cu, which permits the formation of highly soluble, chemically stable products that resist interaction with antagonists in the gut (Brown and Zeringue, 1994). Other studies (Ward et al., 1993) have shown that Cu availability from Cu-lysine and CuSO4 were similar in cattle. Kegley and Spears (1994) also reported that the Cu status of calves fed CuSO4 did not differ from calves fed Cu-lysine. The organic Cu source used in this study (Availa-Cu) is derived from a manufacturing process that produces a Cu-amino acid complex in a metal:amino acid complex ratio of 1:1. Unlike Cu-lysine, where a single amino acid is used, 17 different free amino acids are available to participate in the formation of Availa-Cu (C. K. Swensen, personal communication). Although the availability of studies comparing Cu-lysine to Availa-Cu are lacking, current knowledge would not suggest that they differ significantly. Nevertheless, in the current study, the availability of organic Cu appeared to be similar to that of CuSO4 (Figure 1Go) and TBCC (Table 3Go) when fed in conjunction with a molasses supplement.

The decrease in hay DMI, realized in Exp. 2, is difficult to explain (Figure 2Go). There have been reports from feedlot studies using high-energy finishing diets suggesting that dietary Cu concentrations of 20 ppm are sufficient to reduce gain and feed intake (Ward and Spears, 1997; Engle and Spears, 2000). In those studies, the authors evaluated both organic (Cu proteinate) and inorganic (CuSO4 and TBCC) Cu at 20 ppm in finishing steers over 84 d. During the growing phase, Cu supplementation did not affect steer performance; however, during the finishing phase, each Cu source reduced ADG, ADFI, and gain:feed compared with a control group receiving no supplemental Cu. In another study, finishing steers receiving supplemental TBCC at two levels (10 and 40 ppm) had greater ADG and ADFI compared with steers receiving no supplemental Cu (Engle et al., 2000). It is unclear why these studies differ. Initial Cu status, as well as dietary concentrations of Cu antagonist (Mo, S, and Fe), are potential factors that may influence how cattle respond to Cu supplementation.

Although in vivo evidence is lacking, some in vitro studies suggest that high dietary concentrations of Cu may potentially depress forage fiber digestibility (Church, 1969). In growing steers dosed with a slow-release CuO bolus, OM digestibility of ground hay was reduced compared with a nonbolused control (Arthington and Brown, 2001). In that study, liver Cu concentrations of the steers receiving the CuO bolus were much greater compared with the steers in Exp. 2 of the present study (640 ± 76.3 and 194 ± 30.6 ppm for steers receiving a CuO bolus and steers receiving organic Cu in the current study, respectively). In contrast, Lopez-Guisa and Satter (1992) reported an increase in the digestion of low-quality forages in Holstein cows supplemented with Cu compared with cows consuming a low-Cu diet.

In combination with our first series of studies (Arthington and Pate, 2002), these results provide further insight on the effect of molasses-derived S on Cu nutrition in beef cattle. In the first series of studies, S was found to be responsible for slowing the accumulation of Cu in molasses-supplemented beef heifers. In the current studies, the use of organic Cu did not appear to be an effective alternative to CuSO4 for improving the availability of Cu in molasses supplements. Increasing the concentration of dietary Cu from 10 to 30 ppm resulted in greater liver Cu concentrations in cattle consuming molasses supplements. Despite these findings, cattle from this study and our previous studies were not considered Cu deficient at any point. It is likely that most beef cattle are provided molasses only during defined periods of winter supplementation. Although these data suggest that Cu absorption may be compromised during this time, these cattle likely replenish tissue Cu reserves during the summer months when molasses is not consumed.


   Implications
Top
 Abstract
 Introduction
 Materials and Methods
 Results and Discussion
 Implications
Literature Cited
 
Previous studies indicate that constituents within molasses antagonize the accumulation of dietary copper into the liver of cattle. The use of an organic copper source failed to overcome this antagonism. Feeding higher levels of copper (>10 ppm in the total diet on a DM basis) in molasses-based supplements might be the simplest solution for ensuring adequate copper absorption.


   Footnotes
 
1 Contribution No. R-09109 from the Florida Agriculture Experiment Station. Back

2 Appreciation is expressed to C. Piacitelli and T. Wood for their technical assistance during the conduct of this experiment. The authors also wish to thank the Zinpro Corporation (Eden Prairie, MN) and Micronutrients Inc. (Indianapolis, IN) for their partial financial support of these studies. Back

Received for publication October 21, 2002. Accepted for publication January 22, 2003.


   Literature Cited
Top
 Abstract
 Introduction
 Materials and Methods
 Results and Discussion
 Implications
 Literature Cited
 


Ammerman, C. B. 1969. Recent developments in cobalt and copper in ruminant nutrition: A review. J. Dairy Sci. 53:1097–1107.

Arthington, J. D., and W. F. Brown. 2001. The effect of copper oxide bolus administration on forage fiber digestibility in growing steers. J. Anim. Sci. 79(Suppl. 1):422. (Abstr.)

Arthington, J. D., and L. R. Corah. 1995. Liver biopsy procedures for determining the trace mineral status in beef cows: Part II (Video, AI 8134). Extension TV, Dept. of Communications. Cooperative Extension Service, Kansas State University, Manhattan.

Arthington, J. D., and F. M. Pate. 2002. Effect of corn- versus molasses-based supplements on trace mineral status in beef heifers. J. Anim. Sci. 80:2787–2791.[Abstract/Free Full Text]

Arthington, J. D., J. E. Rechcigl, G. P. Yost, L. R. McDowell, and M. D. Fanning. 2002. Effect of ammonium sulfate fertilization on bahiagrass quality and copper metabolism in grazing beef cattle. J. Anim. Sci. 80:2057–2512.

Braselton, W. E., K. J. Stuart, T. P. Mullaney, and T. H. Herdt. 1997. Biopsy mineral analysis by inductively coupled plasma-atomic emission spectroscopy with ultrasonic nebulization. J. Vet. Diagn. Invest. 9:395–400.[Abstract/Free Full Text]

Brown, T. F., and L. K. Zeringue. 1994. Laboratory evaluations of solubility and structural integrity of complexed and chelated trace mineral supplements. J. Dairy Sci. 77:181–189.[Abstract]

Church, D. C. 1969. Digestive Physiology and Nutrition of Ruminants. Vol. 1. Oregon State University, Corvallis.

Demetriou, J. A., P. A. Drewes, and J. B. Gin. 1974. Ceruloplasmin. Page 857 in Clinical Chemistry. D. C. Cannon and J. W. Winkelman, ed. Harper and Row, Hagerstown, MD.

Engle, T. E., and J. W. Spears. 2000. Effects of dietary copper concentration and source on performance and copper status of growing and finishing steers. J. Anim. Sci. 78:2446–2451.[Abstract/Free Full Text]

Engle, T. E., J. W. Spears, L. Xi., and F. W. Edens. 2000. Dietary copper effects on lipid metabolism and circulating catecholamine concentrations in finishing steers. J. Anim. Sci. 78:2737–2744.[Abstract/Free Full Text]

FASS. 1999. Guide for the Care and Use of Agricultural Animals in Agricultural Research and Teaching. 1st rev. ed. Federation of Animal Science Societies, Savoy, IL.

Kegley, E. B., and J. W. Spears. 1994. Bioavailability of feed-grade copper sources (oxide, sulfate, or lysine) in growing cattle. J. Anim. Sci. 72:2728–2734.[Abstract]

Lopez-Guisa, J. M., and L. D. Satter. 1992. Effect of copper and cobalt addition on digestion and growth in heifers fed diets containing alfalfa silage or corn crop residue. J. Dairy Sci. 75:247–256.[Abstract]

Mason, J. 1981. Molybdenum-copper antagonism in ruminants: A review of the biochemical basis. Irish Vet. J. 35:221–229.

Mason, J. 1990. The biochemical pathogenesis of molybdenum-induced copper deficiency syndromes in ruminants: Towards the final chapter. Irish Vet. J. 43:18–22.

Nockels, C. F., J. DeBonis, and J. Torrent. 1993. Stress induction affects copper and zinc balance in calves fed organic and inorganic copper and zinc sources. J. Anim. Sci. 71:2539–2545.[Abstract]

Rabiansky, P. A., L. R. McDowell, J. Velasquez-Pereira, N. S. Wilkinson, S. S. Percival, F. G. Martin, D. B. Bates, A. B. Johnson, T. R. Batra, and E. Salgado-Madriz. 1999. Evaluating copper lysine and copper sulfate sources for heifers. J. Dairy Sci. 82:2642–2650.[Abstract]

Sumner, S., W. Wade, J. Selph, J. Southwell, V. Hoge, P. Hogue, E. Jennings, P. Miller, and T. Seawright. 1991. Fertilization of established bahiagrass pasture in Florida. Coop. Ext. Service Circular 916, Univ. of Florida.

Suttle, N. F. 1974. Effects of organic and inorganic sulphur on the availability of dietary copper to sheep. Br. J. Nutr. 32:559–568.[Medline]

Ward, J. D., and J. W. Spears. 1997. Long-term effects of consumption of low-copper diets with or without supplemental molybdenum on copper status, performance, and carcass characteristics of cattle. J. Anim. Sci. 73:3057–3065.

Ward, J. D., J. W. Spears, and E. B. Kegley. 1993. Effect of copper level and source (copper lysine vs copper sulfate) on copper status, performance, and immune response in growing steers fed diets with or without supplemental molybdenum and sulfur. J. Anim. Sci. 71:2748–2755.[Abstract]


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