ArticlePDF Available

Effects of Copper and Zinc Supplementation on Weight Gain and Hematological Parameters in Pre-weaning Calves

Authors:

Abstract

Cow-calf operations may be affected by trace mineral deficiencies, particularly copper (Cu) and zinc (Zn) deficiency, which may decrease the calf daily weight gain and alter hematological parameters. We evaluated the effect of Cu and Zn supplementation on pre-weaning calves (n = 40; 92 ± 6 kg initial body weight) from the Salado River basin, Buenos Aires, Argentina. Calves were divided into four groups (n = 10 each) and subcutaneously administered 0.3 mg/kg Cu (Cu group), 1 mg/kg Zn (Zn group), Cu and Zn together (Cu + Zn group), and sterile saline solution (control group) every 40 days for 120 days. Plasma Cu and Zn concentrations, hematological parameters, and weight were recorded every 40 days. A completely randomized 2 × 2 factorial treatment design was used and data were analyzed with a mixed model for repeated measures over time. Cu and Zn were detected in plasma after the second sampling. Cu × Zn interaction was significant (p = 0.09), being Cu concentration higher in the Cu + Zn than in the Cu group. Differences in weight gain (Zn × time interaction; p < 0.01) were observed in the Zn but not in the Cu group (p > 0.1). On the other hand, none of the treatments altered any of the hematological parameters assessed (p > 0.1). Our results show the risk of lower weight gain due to Zn deficiency in pre-weaning calves raised in the Salado River basin.
Effects of Copper and Zinc Supplementation on Weight Gain
and Hematological Parameters in Pre-weaning Calves
Guillermo Alberto Mattioli
1
&Diana Esther Rosa
1
&Esteban Turic
2
&Alejandro Enrique Relling
1
&Esteban Galarza
1
&
Luis Emilio Fazzio
1
Received: 5 December 2017 /Accepted: 29 December 2017
#Springer Science+Business Media, LLC, part of Springer Nature 2018
Abstract
Cow-calf operations may be affected by trace mineral deficiencies, particularly copper (Cu) and zinc (Zn) deficiency, which may
decrease the calf daily weight gain and alter hematological parameters. We evaluated the effect of Cu and Zn supplementation on
pre-weaning calves (n= 40; 92 ± 6 kg initial body weight) from the Salado River basin, Buenos Aires, Argentina. Calves were
divided into four groups (n= 10 each) and subcutaneously administered 0.3 mg/kg Cu (Cu group), 1 mg/kg Zn (Zn group), Cu
and Zn together (Cu + Zn group), and sterile saline solution (control group) every 40 days for 120 days. Plasma Cu and Zn
concentrations, hematological parameters, and weight were recorded every 40 days. A completely randomized 2 × 2 factorial
treatment design was used and data were analyzed with a mixed model for repeated measures over time. Cu and Zn were detected
in plasma after the second sampling. Cu × Zn interaction was significant (p= 0.09), being Cu concentration higher in the Cu + Zn
than in the Cu group. Differences in weight gain (Zn × time interaction; p< 0.01) were observed in the Zn but not in the Cu group
(p> 0.1). On the other hand, none of the treatments altered any of the hematological parameters assessed (p>0.1).Our results
show the risk of lower weight gain due to Zn deficiency in pre-weaning calves raised in the Salado River basin.
Keywords Copper .Zinc .Deficiency .Weight gain .Hematological parameters
Introduction
Trace minerals provide the essential nutrients animals need for
physiological functions, such as growth and development, im-
munity, and reproduction. Consequently, their deficiency can
negatively affect animal performance [1,2]. Copper (Cu) defi-
ciency is the second most frequent mineral deficiency in grazing
cattle worldwide, causing considerable production losses in well-
characterized areas [3]. On the other hand, zinc (Zn) deficiency is
involved in health problems associated with the immune system
and reproductive losses, as well as growth and integrity of the
skin and hooves; however, the pathogenesis remains poorly un-
derstood [3].
In the province of Buenos Aires, Argentina, beef cattle
production represents the main economic activity of the
Salado River basin (SRB). This area covers 5.5 million hect-
ares and produces two million calves a year [4]. Animals are
raised under an extensive system based on naturalized grass as
the main source of nutrients. The economic benefit of the
region resides on selling calves weaned at 67monthsofage.
Different authors have reported Cu and Zn deficiency in the
SRB [57], together with related effects such as decreased
dailyweightgain[3,8,9] and hematological changes [10].
Although the diagnosis of both deficiencies in the herd is
based on the assessment of plasma Cu and Zn concentrations,
there are discrepancies regarding data interpretation. In terms
of Cu, concentrations > 57 μg/dL are considered adequate and
<57 μg/dL indicate a deficiency [3]. Nevertheless, these au-
thors reported clinical symptoms of hypocuprosis with Cu
levels < 30 μg/dL [3]. In contrast, Cu concentrations of 50
70, 2050, and < 20 μg/dL are considered as marginal, defi-
cient, and clinically evident disease, respectively [11].
Regarding Zn, concentrations > 90 μg/dL are considered to
be adequate, 8090 μg/dL as marginal, and < 80 μg/dL as
deficient [2]. Both Cu and Zn deficiencies are associated with
*Guillermo Alberto Mattioli
mattioli@fcv.unlp.edu.ar
1
Laboratorio de Nutrición Mineral, Facultad de Ciencias Veterinarias,
Universidad Nacional de La Plata, 60 y 118, 1900 La
Plata, Argentina
2
Biogenésis Bagó, Buenos Aires, Argentina
Biological Trace Element Research
https://doi.org/10.1007/s12011-017-1239-0
hematological changes such as anemia, leukopenia, and al-
tered tissue enzymes [10,12].
Here we discuss whether plasma Cu and Zn concentrations
are increased after parenteral Cu, Zn, and Cu + Zn supplemen-
tation of pre-weaning calves, thereby altering daily weight
gain and hematological parameters.
Materials and Methods
All experimental procedures were approved by the Committee
for the Care and Use of Laboratory Animals (CICUAL, for its
Spanish acronym), School of Veterinary Sciences, La Plata
National University, Argentina (Protocol no. 58-2-16P).
The trial was carried out on the experimental farm
BManantiales,^located in Chascomús, Buenos Aires (35°
4431.5^S58°0611.7^W). The characteristics of the farm
are comparable to those in the SRB, including poor drainage,
floods, and higher quantity and quality grass production dur-
ing spring.
Animals
A total of 40 clinically healthy Aberdeen Angus calves were
used. They were kept as cow-calf pairs since time 0 of the trial
(3 months of age) until weaning (7 months of age; time 120 of
the trial). The vaccination program of calves included foot-
and-mouth disease vaccine and two doses of clostridial vac-
cine before weaning. Gastrointestinal parasites were monthly
examined through fecal egg counts.
The animals were fed on native and naturalized grass
(Chaetotropis elonga,Stenotaphrum secundatum,Paspalum
dilatatum,Lolium perenne,Lotus tenuis)withwidegrass
availability and ad libitum water consumption.
Groups and Treatments
Calves were assigned to one of four homogeneous groups
according to weight, sex, and age (n= 10 each group), and
treated as follows: Cu group, 0.3 mg/kg Cu edetate; Zn group,
1 mg/kg Zn edetate; Cu + Zn group, the same doses of Cu and
Zn edetate (Suplenut®, Biogénesis Bagó); and control group,
saline sterile solution. The animals were subcutaneously
injected every 40 days from November 2015 to March 2016
on days 0, 40, 80, and 120 days.
Blood samples were obtained via jugular venipuncture and
collected in Na
2
EDTA-containing tubes previously washed
with deionized water. They were kept at 4 °C until processing
within 6 h after collection. Blood was centrifuged at 1500 rpm
for 10 min and plasma was proportionally deproteinized with
10% trichloroacetic acid. Copper and Zn concentrations were
measured in supernatants using atomic absorption spectropho-
tometry (AAS) (Perkin Elmer AAnalyst 200). Blood samples
(n= 5 per group) were collected in tubes containing EDTA-K
3
and sent to a regional laboratory (Laboratorio Azul SA) to be
processed for evaluation of hematological parameters (red cell
count, hemoglobin concentration, hematocrit percentage,
hematimetric indices, white blood cell count, absolute and
relative leukocyte formula, and platelets count).
At the same time, grass was collected from three sites in the
paddock according to animal behavior and forage intake.
Samples were washed, dried, and exposed to acid digestion
(3:1 nitric-perchloric acid mixture). The concentrations of Cu,
Zn, and iron (Fe) were measured with AAS, whereas molyb-
denum (Mo) and sulfur (S) concentrations were measured
using graphite furnace AAS and Arsenazo III titration
(adapted from Hamm et al. [13]), respectively. In grass sam-
ples, Zn, Cu, Mo, and Fe concentrations were measured.
The quality of bovine drinking water was analyzed in a
sample initially collected from water troughs, the only water
source for the animals.
Individual animal weight was recorded early in the morn-
ing after the animals had fasted for 12 h.
Statistical Design and Analysis
We used a completely randomized statistical design. Data
were analyzed using a mixed model of repeated measures over
time and a 2 × 2 factorial arrangement using SAS statistical
software 9.1. The main factor was parenteral supplementation
with or without Zn or Cu. Supplementation with Zn and/or
Cu, all their possible interactions and time were taken as fixed
variables, whereas the animals represented the random vari-
able. The SLICE option was used for mean separation if sig-
nificant differences were reported for the main variables
(p< 0.05), interactions (p< 0.1), or tendencies (p<0.15).
When only the treatment was significant, mean separation
was done by a protected Fishers test using the PDIFF-SAS
option. Associations between plasma Cu and Zn concentra-
tions and weight gain were assessed with correlation analysis
using the same statistical software.
Results
In the Cu group, plasma Cu concentration increased after the
second sampling (Cu × time interaction, p<0.01; Table 1).
We also found Cu × Zn interaction (p=0.09;Table 1), being
plasma Cu concentration higher in the Cu + Zn than in the Cu
group (76.2 vs. 73.4 ± 2.1 μg/dL). In turn, plasma Cu concen-
tration was lower in the Zn than in the control group (46.2 vs.
50.7 ± 2.1 μg/dL). In the case of plasma Zn concentration, it
Mattioli et al.
increased after Zn supplementation either alone or together
with Cu (p=0.02; Table1).
In terms of weight, time differences were observed in the
Zn-treated group, finding higher weight gain after the second
sampling (p<0.02; Table 1). Both Cu and Zn concentration
correlated with weight (Cu: r=0.04, p=0.53; Zn: r=0.25,
p<0.01).
Regarding hematological parameters (erythrocytes, leuko-
cytes, and enzymes), no differences were detected in any of
the four study groups (Table 2).
In grass samples, Zn, Cu, Mo, and Fe concentrations were
19±5, 7.3±1.2, 0.7±0.4, 329±80 ppm dry matter (DM),
respectively, and S concentration was 0.13 ± 0.06% DM. No
significant Cu and Zn contents were detected in drinking
water.
Discussion
Cu deficiency may result in reduced daily weight gains in
calves [3], particularly in cases of severe deficiency [11]. In
the present study, marginal Cu deficiency did not affect either
daily weight gain or hematological parameters. Nevertheless,
our data correlated with previous results of Cu supplementa-
tion at the SRB reporting differences in weight gain in calves
with Cu levels lower than 25 μg/dL [14], which are related to
severe deficiency [15]. Likewise, decreased hemoglobin con-
centrations and low white cell counts were found in heifers
only with Cu concentrations lower than 19 μg/dL [12]. The
marginal Custatus reported in the present trial would be due to
the Cu concentration in grass (7.3 ppm DM), which was lower
than the required 10 ppm DM [16]. Moreover, other grass-
related factors that may lead to Cu deficiency, such as Mo,
Fe, and S, showed a moderate concentration, suggesting that
they did not interfere with Cu absorption [17].
AninterestingfindingrelatedtoCubehaviorwastheCu×Zn
interaction (p= 0.09) in the Cu + Zn group, since supplementation
produced higher Cu concentrations as compared with the Cu
group. Probably, Zn could have promoted metallothionein syn-
thesis in the liver with the combined supplementation [18]. This
protein enhances Cu capture and acts as a liver Cu storage through
which ceruloplasmin is produced to constitute the main determi-
nant of plasma Cu concentration [1,19,20]. Furthermore, Cu
concentrations in the Zn group were lower than in the control
group. Supplementation with Zn could have increased Cu require-
ments since some mechanisms depend on both elements. For
example, Cu-Zn superoxide dismutase (Cu-Zn SOD) is an en-
zyme whose action is related to Zn concentration and also requires
Cu for an adequate activity [21].
The study groups supplemented with Zn either alone or
combined with Cu presented higher weight regardless of
Cu supplementation, probably due to the altered intake
and/or feed conversion produced by Zn deficiency in bo-
vines [2,8]. Studies on experimental animals indicate that
Zn deficiency leads to lower water intake [22], altered
thyroid function [23], IGF-1 signaling failure [24], and
Table 1 Copper (Cu) and zinc (Zn) concentration least square means and live weight of pre-weaning calves in the four study groups
Day Groups SEM pvalue
1
Control Cu Zn Cu + Zn Zn Cu Zn × Cu Zn × T Cu × T
Cu concentration (μg/dL)
0 46 42 44 45 1.5 0.67 < 0.01 0.09 0.96 < 0.01
40 49 88 45 89
80 53 87 49 89
120 55 76 47 81
Zn concentration (μg/dL)
0 84 80 85 81 2.3 0.02 0.12 0.77 0.48 0.98
40 112 101 115 117
80 108 102 115 109
120 92 91 109 100
Weight (kg)
0 92 92 93 92 1.0 0.02 0.73 0.43 < 0.01 0.48
40 124
a
124
a
127
b
126
b
80 145
a
146
a
148
b
149
b
120 172
a
173
a
181
b
177
b
SEM standard error of the mean, Ttime
Different letters in the same row indicate p<0.05
1
There was no Cu × Zn × time interaction for any of the variables
Effects of Copper and Zinc Supplementation on Weight Gain and Hematological Parameters in Pre-weaning Calves
anorexia secondary to the suppression of hypothalamic
neuropeptide Y [25].
The National Research Council recommends 30 ppm DM
of Zn to reach the requirements [16] and suggests that lower
average weight gain could occur with Zn dietary doses of
20 ppm DM [11,26]. These data are in agreement with the
19 ppm DM Zn found in the present trial; Zn concentration
was higher in the Zn groups and correlated with weight gain
(r:0.25;p< 0.01). Although it is agreed that Zn concentration
should be taken as an indicator of Zn status in animals, most of
the trials reporting differences in terms of weight gain in
groups supplemented with or without Zn showed similar Zn
concentrations [27,28]. The time (weeks-months) required for
the development of the deficiency as an indicator of Zn status
might improve Zn concentration. In a previous trial, it was
reported that 6 weeks were needed to distinguish the Zn-
supplemented (40 ppm DM) from the control group
(17 ppm) [29]. Other studies obtained similar results in
3 weeks, i.e., low daily weight gain but no differences in
plasma Zn concentration [8]. In this trial, Zn supplementation
every 40 days during 4 months was associated with higher
weight gain. Further research showing the importance of herd
risk diagnosis based on plasma Zn concentration could con-
tribute to preventing Zn deficiency in calves.
Conclusions
Zinc parenteral supplementation every 40 days improved the
daily weight gain of calves, indicating the risk for Zn deficien-
cy in the SRB area. Marginal Cu concentration did not induce
lower weight gain, and marginal Cu + Zn concentration did
not alter hematological parameters.
Acknowledgements The authors would like to thank A. Di Maggio for
manuscript correction and edition and to Dr. Darío Piacentini for edetate
Cu and Zn solutions.
Author Contributions G.A.M. and L.E.F. conceived and designed the
experiments; D.E.R., and E.M.G. conducted the experiments; A.E.R.
analyzed the data; E.T., G.A.M., and L.E.F. critically wrote and revised
the paper. All authors read and approved the final manuscript.
Funding This study was supported by a grant from the National Program
of Incentives to Teaching and Research, Secretary of University Policies,
Ministry of Education of Argentina (grant no. 11/V204, School of
Veterinary Sciences, National University of La Plata). The sponsor had
no involvement in the study design, collection, analysis, or interpretation
of the data presented in this paper.
Compliance with Ethical Standards
Conflict of Interest The authors declare that they have no competing
interests.
References
1. Richards MP (1989) Recent developments in trace element metab-
olism and function: role of metallothionein in copper and zinc me-
tabolism. J Nutr 119(7):10621070
Table 2 Least square means for hematological and serological
parameters in pre-weaning calves from the four study groups
Day Groups*
Control Cu Zn Cu + Zn
Red cell/mm
3
0 6442 6566 6438 6540
40 6538 6444 6540 6652
80 7618 7286 6988 7482
120 6978 6746 6880 6752
Hemoglobin (g/dL)
0 13.08 13.32 13.00 13.22
40 13.18 13.26 13.14 13.68
80 13.90 13.36 13.46 14.08
120 13.18 13.46 13.66 13.28
Hematocrit (%)
040434144
40 41 41 41 41
80 45 43 42 44
12042424442
Leukocytes/mm
3
0 6680 4645 5437 6108
40 5000 5820 5100 6200
80 5720 6320 7260 5920
120 5780 6040 5840 5780
ALP (U/L)
0 159 165 154 146
40 165 165 159 200
80 178 180 190 174
120 201 172 205 172
AST (U/L)
0 93 94 113 94
40 92 104 89 101
80 117 107 117 102
120 98 109 111 102
GGT (U/L)
026313134
40 34 34 29 30
80 35 33 29 29
12029313033
SEM standard error of the mean, ALP serum alkaline phosphatase (units
per liter), AST aspartate aminotransferase (units per liter), GGT gamma
glutamyl transpeptidase (units per liter)
*
There was no Cu × Zn × time interaction for any of the variables
Mattioli et al.
2. Enjalbert F, Lebreton P, Salat O (2006) Effects of copper, zinc and
selenium status on performance and health in commercial dairy and
beef herds: retrospective study. J Anim Physiol Anim Nutr (Berl)
90(1112):459466. https://doi.org/10.1111/j.1439-0396.2006.
00627.x
3. Suttle NF (2010) Copper. In: Suttle NF (ed) The mineral nutrition of
livestock, 4th edn. Oxfordshire, UK, pp 255305. https://doi.org/
10.1079/9781845934729.0255
4. Némoz JP (2013) Principales características de la actividad
ganadera vacuna en el área de la EEA Cuenca del Salado.
Informe INTA-RIAN. Available at https://inta.gob.ar/documentos/
principales-caracteristicas-de-la-actividad-ganadera-vacuna-en-el-
area-de-la-eea-cuenca-del-salado. Accessed 15 Nov 2017
5. Mattioli GA, Ramírez CE, Giuliodori MJ, Tittarelli CM, Yano H,
Matsui H (1996) Characterization of cattle copper deficiency in the
Magdalena District. Livest Prod Sci 47(1):710. https://doi.org/10.
1016/S0301-6226(96)01004-4
6. Ramírez CE, Mattioli GA, Tittarelli CM, Giuliodori MJ, Yano H
(1998) Cattle hypocuprosis in Argentina associated with periodical-
ly flooded soils. Livest Prod Sci 55(1):4752. https://doi.org/10.
1016/S0301-6226(98)00120-1
7. Ramírez CE, Mattioli GA, Giuliodori MJ, Yano H, Matsui I (1998)
Deficiencia de Zn en bovinos de cría de la provincia de Buenos
Aires. Vet Argent 15(142):114118
8. Engle TE, Nockels CF, Kimberling CV, Weaber DL, Johnson AB
(1997) Zinc repletion with organic or inorganic forms of zinc and
protein turnover in marginally zinc-deficient calves. J Anim Sci
75(11):30743081. https://doi.org/10.2527/1997.75113074x
9. Arrayet JL, Oberbauer AM, Famula TR, Garnett I, Oltjen JW,
Imhoof J, Kehrli ME, Graham TW (2002) Growth of Holstein
calves from birth to 90 days: the influence of dietary zinc and
BLAD status. J Anim Sci 80(3):545552. https://doi.org/10.2527/
2002.803545x
10. Sharma MC, Joshi C (2005) Therapeutic efficacy of zinc sulphate
used in clustered model treatment in alleviating zinc deficiency in
cattle and its effect on hormones, vitamins and production parame-
ters. Vet Res Commun 29(7):609628. https://doi.org/10.1007/
s11259-005-3382-x
11. Kincaid RL (1999) Assessment of trace mineral status of ruminants:
a review. Proc Am Soc Anim Sci 77(1):110
12. Sharma MC, Joshi C, Pathak NN, Kaur H (2005) Copper status and
enzyme, hormone, vitamin and immune function in heifers. Res Vet
Sci 79(2):113123. https://doi.org/10.1016/j.rvsc.2004.11.015
13. Hamm JW, Bettany JR, Halstead EH (1973) A soil test for sulfur
and interpretative criteria for Saskatchewan. Comm Soil Sci Plant
Anal 4:219231
14. Fazzio LE, Mattioli GA, Picco SJ, Rosa DE, Minatel L, Gimeno EJ
(2010) Diagnostic value of copper parameters to predict growth of
suckling calves grazing native range in Argentina. Pesquisa
Veterinária Brasileira 30(10):827832. https://doi.org/10.1590/
S0100-736X2010001000004
15. Dargatz DA, Garry FB, Clark GB, Ross PF (1999) Serum copper
concentrations in beef cows and heifers. J Am Vet Med Assoc
215(12):18281832
16. National Research Council (2000) Nutrient requirements of beef
cattle, seventh revised edition: update 2000. The National
Academies Press, Washington, DC, pp 5474. Available at https://
www.nap.edu/catalog/9791/nutrient-requirements-of-beef-cattle-
seventh-revised-edition-update-2000. Accessed 15 Nov 2017
17. Gould L, Kendall NR (2011) Role of the rumen in copper and
thiomolybdate absorption. Nutr Res Rev 24(2):176182. https://
doi.org/10.1017/S0954422411000059
18. Formigari A, Irato P, Santon A (2007) Zinc, antioxidant systems
and metallothionein in metal mediated-apoptosis: biochemical and
cytochemical aspects. Comp Biochem Physiol C Toxicol
Pharmacol 146(4):443459. https://doi.org/10.1016/j.cbpc.2007.
07.010
19. Arredondo M, Núñez MT (2005) Ironand copper metabolism. Mol
Asp Med 26(45):313327. https://doi.org/10.1016/j.mam.2005.
07.010
20. Liu J, Zhou ZX, Zhang W, Bell MW, Waalkes MP (2009) Changes
in hepatic gene expression in response to hepatoprotective levels of
zinc. Liver Int 29(8):12221229. https://doi.org/10.1111/j.1478-
3231.2009.02007.x
21. Bengoumi M, Essamadi K, Charcornac JP, Tressol JC, Faye B
(1998) Comparative relationship between copper-zinc plasma con-
centrations and superoxide dismutase activity in camels and cows.
Vet Res 29(6):557565
22. Norii T (2008) Growth of zinc-deficient rats during intra-gastric
tube feeding. Biol Trace Elem Res 122(3):266275. https://doi.
org/10.1007/s12011-007-8073-8
23. Jing MY, Sun JY, Wang JF (2008) The effect of peripheral admin-
istration of zinc on food intake in rats fed Zn-adequate or Zn-
deficient diets. Biol Trace Elem Res 124(2):144156. https://doi.
org/10.1007/s12011-008-8132-9
24. MacDonald RS (2000) The role of zinc in growth and cell prolifer-
ation. J Nutr 130(5):15001508
25. Levenson CW (2003) Zinc regulation of food intake: new insights
on the role of neuropeptide Y. Nutr Rev 61(7):247249
26. Mufarrege DJ, Aguilar DE (2001) Suplementación con zinc de los
bovinos para carne en la provincia de Corrientes. E.E.A. INTA
Mercedes, Corrientes, Noticias y Comentarios Nº 348: 14.
Available at http://www.produccion-animal.com.ar/
suplementacion_mineral/58-suplementacion_con_zinc.pdf.
Accessed 15 Nov 2017
27. Beeson WM, Perry TW, Zurcher TD (1977) Effect of supplemental
zinc on growth and on hair and blood serum levels of beef cattle. J
Anim Sci 45(1):160165. https://doi.org/10.2527/jas1977.451160x
28. Spears JW, Kegley EB (2002) Effect of zinc source (zinc oxide vs
zinc proteinate) and level on performance, carcass characteristics,
and immune response of growing and finishing steers. J Anim Sci
80(10):27472752. https://doi.org/10.2527/2002.80102747x
29. Neathery MW, Miller WP, Blackmon DM, Gentry RP, Jones JB
(1973) Absorption and tissue zinc content in lactating dairy cows
as affected by low dietary zinc. J Anim Sci 37(3):848852. https://
doi.org/10.2527/jas1973.373848x
Effects of Copper and Zinc Supplementation on Weight Gain and Hematological Parameters in Pre-weaning Calves
... I mproving animal health and production requires careful attention to nutritional status and nutrition composition, which is not only concerned with protein, carbohydrates, and major elements but it is dependent on trace minerals. Trace minerals are essential for growth and development, immunity, and reproduction; they act as a co-factor in the physiological functions of cells, cell metabolism, and the oxidation process (Mattioli et al., 2018). ...
... The current work showed insignificant changes in the hemogram between groups except for a significant increment in MCV and a significant decrement in MCHC at D30. Yanagisawa et al. (2009) reported that rats administered a high Zn diet showed reticulocytosis and extra-medullary erythropoiesis in the spleen which could be responsible for the maintenance of RBC levels. Mattioli et al. (2018) found no change in the assessed hematological parameters in pre-weaning calves supplemented with Cu and Zn. Moreover, Cope et al. (2009) denoted that erythrocytes and leukocytes were unaffected by the dietary zinc levels. ...
... In our study, we observed reduced intake of mineral supplement at higher doses of monensin, in line with previous studies (Fitzgerald and Mansfield 1973), which may have prevented the weight gain expected at the full dose. The effect of mineral supplementation on weight gain has been observed by others as well (Dias et al. 2013;Gunter and Combs 2019;Mattioli et al. 2018). ...
Article
Full-text available
Monensin is known to improve feed efficiency in cattle. At higher doses, however, monensin reduces the palatability of mineral supplements, which may reduce consumption of essential minerals such as selenium. The main objective of this study was to compare weight gain, while evaluating the impact on blood selenium concentrations, among treatment and control groups of stocker calves supplemented with a self-fed mineral supplement designed to deliver different amounts of monensin, while the control group received the same mineral without monensin. A secondary objective was to compare the presence of pathogenic and total coccidia oocysts. At study end, all monensin treatment groups weighed more than the control group but were not different from each other. No effect was detected on coccidia oocysts. In sum, there is a production gain from providing monensin in a mineral supplement to weaned calves on pasture. However, too high a dose may lead to insufficient trace mineral consumption. This is a concern in areas where forage is deficient in these nutrients.
... Different studies showed that nutritional interventions during the gestation period improve the immune response and milk/colostrum quality and reduce oxidative damage [23,24]. The studies have reported that micronutrient supplementation such as selenium, manganese, zinc, and vitamins A, C, and E improved the maternal vaccination responses, immune functions, and antibody production, and reduced stress levels and maternal hyperglycemia during pregnancy and parturition [24][25][26][27][28]. Recent research has suggested that micronutrient supplements with COVID-19 vaccines positively correlated with immunoglobulin levels [29][30][31]. ...
Article
Full-text available
Contagious ecthyma is a contagious zoonotic disease caused by the Orf virus that can infect farm animals and humans, but no vaccine is available for pregnant mothers. Excessive oxidative stress during pregnancy can suppress the vaccine immune response in pregnant mothers; hence, maternal micronutrient supplementation could effectively improve the immune response, health, and oxidative status during pregnancy. In this study, we employed an 8-week-old pregnant rat model to receive a single intramuscular dose of 200 µg of ORF DNA vaccine with or without vitamin E and selenium supplementation to evaluate their effect on immune responses (specific IgG and IgG isotypes), oxidative stress, liver enzymes, and blood glucose levels in maternal-neonatal serum and milk secretions. Additionally, antioxidant-related gene expressions were analyzed in the maternal placenta and pups’ liver. The results showed that supplementation of vitamin E and selenium with ORF DNA vaccination increased the production of specific antibody and IgG isotypes (IgG1 and IgG2a) and reduced the oxidative stress in neonatal-maternal serum and milk compared to both the control group and those vaccinated without supplementation (p < 0.05). Notably, the ORF DNA vaccine did not cause oxidative stress and hepatic damage. However, combined supplementation of vitamin E and selenium with DNA vaccination significantly decreased serum malondialdehyde (MDA) levels and improved the antioxidant-related enzyme activities of glutathione peroxidase (GPX), superoxide dismutase 1 (SOD1), and selenoprotein P (SELP) in the maternal placenta and liver of pups (p < 0.05). In conclusion, maternal supplementation of vitamin E and selenium enhanced the immune responses of the ORF DNA vaccine by mitigating oxidative stress in pregnant rats and could thus be a promising strategy for better health outcomes for both mothers and neonates.
... Western blot samples were also used to assess intracellular Cu concentration, following established procedures [32]. In brief, the samples underwent acid digestion using a 3:1 mixture of nitric and hydrochloric acid. ...
Article
Full-text available
Background Cholesterol (Cho) is an essential lipophilic molecule in cells; however, both its decrease and its increase may favor the development of neurological diseases such as Alzheimer’s disease (AD). Although copper (Cu) is an essential trace metal for cells, the increased plasma concentration of its free form has been linked with AD development and severity. AD affects aged people, but its prevalence and severity are higher in women than in men. We have previously shown that Cu promotes Cho de novo synthesis in immature neurons as well as increased Cho in membrane rafts and Aβ levels in culture medium, but there are no results yet regarding sex differences in the effects of sublethal Cu exposure on Cho de novo synthesis. Methods We examined the potential sex-specific impact of sublethal Cu concentrations on de novo Cho synthesis in primary cultures of male and female astrocytes. We also explored whether this had any correlation with variations in Cho and APP levels within neuronal membrane rafts. Results Flow cytometry analysis demonstrated that Cu treatment leads to a greater increase in ROS levels in female astrocytes than in males. Furthermore, through RT-PCR analysis, we observed an upregulation of SREBP-2 and HMGCR. Consistently, we observed an increase in de novo Cho synthesis. Finally, western blot analysis indicated that the levels of ABCA1 increase after Cu treatment, accompanied by a higher release of radiolabeled Cho and an elevation in Cho and APP levels in neuronal membrane rafts. Importantly, all these results were significantly more pronounced in female astrocytes than in males. Conclusions Our findings confirm that Cu stimulates Cho synthesis in astrocytes, both in a ROS-dependent and -independent manner. Moreover, female astrocytes displayed elevated levels of HMGCR, and de novo Cho synthesis compared to males following TBH and Cu treatments. This corresponds with higher levels of Cho released into the culture medium and a more significant Cho and APP rise within neuronal rafts. We consider that the increased risk of AD in females partly arises from sex-specific responses to metals and/or exogenous substances, impacting key enzyme regulation in various biochemical pathways, including HMGCR.
... Additionally, because the immune system of neonatal calves is still developing, they are more susceptible to infectious pathogens. Therefore, calves rely on the effective passive transfer of maternal Ig from the colostrum after birth [22][23][24][25][26]. Trace elements, which play significant roles in nutrition and regulate many critical biological processes, are important for optimizing the production of beef cattle [27][28][29][30][31]. Trace mineral deficiencies or impaired placental transfer of these minerals during fetal life can adversely affect not only overall growth but also the immunological and morphological development of a variety of fetal and neonatal tissues [32,33]. ...
Article
Full-text available
Simple Summary Beef calves are born with underdeveloped immune systems and extremely vulnerable to diseases. Therefore, calf health is one of the most significant animal health issues facing the livestock industry. Maternal nutrition during pre- and post-partum with essential nutrients plays a significant role in offspring’s physiological functions. Thus, this study attempted to evaluate the effects of supplementation of the maternal diet with organic trace minerals on the health and immune status of beef calves. This study indicated that maternal supplementation with trace minerals containing zinc, manganese, copper, and cobalt is a promising strategy for preventing infections and improving calves’ immunity. Abstract In this study, we evaluated the effects of supplementation of the maternal diet with organic trace minerals including Zn (zinc), Mn (manganese), Cu (copper), and Co (cobalt) on the health and immune status of beef calves. We examined 19 pregnant cows, which were divided into a group of 9 cows fed a basal diet (control) and 10 cows fed a diet with organic trace minerals (treated). Cows were fed for a period of 45 days before the predicted calving date until 45 days after calving. The number of treatments needed for respiratory and digestive diseases within 14 days of birth was significantly lower in the treated group (p < 0.05) than the control group. In addition, the concentration of serum zinc in the treated group on day 1 was significantly higher (p < 0.05) than that in the control group. The numbers of CD4⁺ and CD8⁺ cells in the treated group on days 30 and 60 were significantly increased (p < 0.01) compared with those in the control group, as was the number of γδ T cells on days 1 and 30 (p < 0.05). The number of IgM⁺ cells in the treated group on days 30 and 60 was significantly increased (p < 0.01) compared with that in the control group, as was the number of MHC class II⁺ cells on day 60 (p < 0.01). The number of NK cells in the treated group on day 60 was also significantly increased (p < 0.05) compared with that in the control group. The expression levels of mRNAs encoding interlukin-2 (IL-2), interlukin-4 (IL-4), interlukin-12 (IL-12), and interferon-γ (IFN-γ) in the treated group were significantly higher than those in the control group (p < 0.05) on days 1 and 60. The results indicate that maternal supplementation with trace minerals is a promising approach for producing highly disease-resistant calves and enhancing calf immunity.
... Ht, MCV, and TPC values showed no difference between groups, and their results are similar to those obtained by [37] in Nellore and Holstein breeds, [48] in Norwegian Red, [49] in Pantaneira, and [9] in Hanwoo; while the variables Hb and CHC showed values compatible with similar studies in other races and different age groups [50][51][52]. Age did not influence the behavior of the erythrometric variables between the groups, corroborating the results of [40,49,53,54]. ...
Article
Full-text available
The Brazilian savannah region, characterized by high average temperatures, well-defined rainy and dry seasons, soil with low productive potential, and high pressure for parasitic diseases, is home to the highest percentage of the beef herd, which is the world’s largest commercial beef producer. Therefore, breeds that present rusticity combined with productivity are the focus of research in cattle breeding in the region. Considering their geographic particularities and their effects on the animals’ blood parameters, the objective was to study the behavior of hematological variables, serum biochemistry, and acute phase proteins in beef calves at weaning age. Jugular blood samples were collected in a single day from 30 weaning calves (about eight months old and 200 kg of body weight) from the following breeds: Nellore, Senepol, Nellore × Aberdeen Angus cross, Nellore × Senepol cross, and Nellore × Aberdeen Angus × Senepol cross. Hematological data were obtained using an automatic cell counter, serum biochemical measurements were obtained using commercial kits, and the electrophoretogram was obtained using the SDS-page technique. In general, the results were consistent with data already published in similar situations regarding health status, age, and level of metabolic activity. However, differences observed between groups can be explained by differences observed in other concurrent variables like temperament. The pure zebu breed was more reactive than pure taurine. While crossbreds showed intermediate values, and parasitic infestation, the pure taurine breed with higher parasite infestation, while the zebu breed had lower values, which produced effects in some laboratory tests, and generated differences between breeds. In conclusion, the reference intervals available for healthy animals can be routinely used without interference from the geographic region for animals produced without nutritional failures as long as changes are recorded in pathological, infectious, metabolic, or nutritional deficiency situations. However, it is suggested that a study covering a larger number of herds may demonstrate a greater geographic effect on the studied variables.
... The amount equivalent to 0.9 mg/day of copper, despite causing changes in some of the regulatory mechanisms of cardiac contractility, was not able to reduce the force generation of papillary muscles. It was observed that the body weight gain during the 4 weeks of exposure was similar among the experimental groups that received 13 µg, 0.14 mg or 0.28 mg/kg/day of copper, corroborating a study by Naseri et al. [32] and Mattiolli et al. [33], in which steers that received copper supplementation up to the maximum recommended level showed no change in body mass gain. To investigate whether the cardiac effects were not due to other factors, such as vascular effects leading to pressure overload, we tested the copper effects on aortic segments. ...
Article
Full-text available
Copper is essential for homeostasis and regulation of body functions, but in excess, it is a cardiovascular risk factor since it increases oxidative stress. The objective of this study was to evaluate the effects of exposure to the recommended daily dose (13 µg/kg/day), upper tolerable dose (0.14 mg/kg/day) and twice the upper tolerable dose (0.28 mg/kg/day) via i.p. over 4 weeks on the vascular reactivity of aortic rings and the contraction of LV papillary muscles of male Wistar rats. It was also determined whether the antioxidant peptide from egg white hydrolysate (EWH) prevents these effects. Copper exposure at the doses evaluated did not change weight gain of male Wistar rats, the reactivity of the aortic rings or the cardiac mass. The dose of 0.13 µg/kg/day did not reduce the force of contraction, but it impaired the time derivatives of force. Doses of 0.14 and 0.28 mg/kg/day reduced the force of contraction, the inotropic response to calcium and isoproterenol, the postrest contraction and the peak and plateau of tetanized contractions. EWH treatment antagonized these effects. These results suggest that copper, even at the dose described as upper tolerable, can impair cardiac contraction without altering vascular reactivity. Antioxidative stress therapy with EWH reversed these harmful effects, suggesting a possible strategy for the amelioration of these effects.
... The health status of newborn calves is one of the most important issues facing the livestock industry, and preservation of their well-being is equally essential as today's calves are known to be tomorrow's herd (Lorenz et al., 2011;Mattioli et al., 2018). Calves are characterized by an underdeveloped immune system at birth, thus, they depend immunologically on the successful passive transfer of maternal immunoglobulins from colostrum after birth (Salmon, 1999). ...
Article
Full-text available
This study aimed to investigate the immune-metabolic status and growth performance of Simmental calves born from cows subjected to pegbovigrastim administration 7 days before calving. Eight calves born from cows subjected to pegbovigrastim administration (PEG group) and 9 calves born from untreated cows (CTR group) were used. Growth measurements and blood samples were collected from birth to 60 d of age. The PEG group had lower body weight from 28 up to 60 d of age (P < 0.01), lower heart girth (P < 0.05), lower weekly and total average daily gain values (P < 0.05) than the CTR group throughout the monitoring period. A decrease in milk replacer (MR) intake was observed in the PEG group compared with the CTR group around 20-28 d of age (P < 0.01). The PEG group had lower values of γ-glutamyl transferase (GGT) at d 1 of age (P < 0.05), Zn at 21 and 28 d of age (P < 0.05), hemoglobin, MCH and MCHC at 54 and 60 d of age (P < 0.01), and higher urea concentration at 21 and 28 d of age (P < 0.05) compared with the CTR group. Lower values of retinol (P < 0.05), tocopherol (P < 0.01), mean myeloperoxidase index (P < 0.05) and higher total reactive oxygen metabolites (P < 0.05) and myeloperoxidase (P < 0.05) were also detected in the PEG group. In light of the results gathered in the current study, it can be speculated that activation of the cow's immune system by pegbovigrastim could have influenced the immune competence, growth performance as well as the balance between oxidant and antioxidant indices of the newborn calf.
... The health status of newborn calves is one of the most important issues facing the livestock industry, and preservation of their well-being is equally essential as today's calves are known to be tomorrow's herd (Lorenz et al., 2011;Mattioli et al., 2018). Calves are characterized by an underdeveloped immune system at birth, thus, they depend immunologically on the successful passive transfer of maternal immunoglobulins from colostrum after birth (Salmon, 1999). ...
Article
Full-text available
A study was conducted to evaluate the predictive diagnostic value of different copper (Cu) parameters as indicators of average daily gain (ADG) in growing calves. The effects in calves of cow Cu supplementation in the last one-third gestation period were also evaluated. Five supplementation trials, with a total of 300 calves, were carried out. Two groups of 30 calves were randomly assigned to each trial, one group was parenterally supplemented (SG) and the other was not supplemented (NSG). Trials began when calves were three-month-old and ended at weaning time. At each sampling calves were weighed and blood was taken to determine Cu concentrations in plasma, Whole Blood (WB), Red Cells (RC) and Packed Cell Volume (PCV). Liver samples from six animals of each group were taken both at the beginning and at the end of the trial. In two trials the mothers of the SG received Cu supplementation at the last one- third gestation period. Four of the five trials exhibited low ADG in the NSGs. In these groups, plasma Cu concentration decreased rapidly before low ADG was detected, which occurred with values remaining below 25µg/dl. The decrease of RC Cu concentration was considerably slow. WB showed an intermediate position. PCV in the SGs was higher than in the NSGs in all trials. Cow supplementation was insufficient to generate a liver storage able to last after calves reached the 3 months of age. These data could be useful to predict the risk of low ADG in grazing calves.
Article
Full-text available
An assessment of the trace mineral status of ruminants begins with clear objectives. These may be to determine whether a nutrient deficiency exists, to assess the prevalence of a deficiency, or to estimate the endogenous reserves of trace minerals. The sampling size needed for assessment is a function of the herd size, expected prevalence of deficiency or toxicity, desired confidence of the assessment tool, and the estimated standard deviation of the assessment criteria. The trace mineral status of animals is best described by concentrations in liver. Correlation coefficients between concentrations of trace minerals in blood and liver are highest in deficient animals because endogenous reserves are depleted. Concentrations of Zn, Cu, and Se in plasma also are affected by infection, stress, pregnancy, and erythrocyte hemolysis. Because erythrocytes in cattle have a 160-d life span, concentrations of trace minerals in whole blood change more slowly than those in plasma in response to changes in intakes of trace minerals. Improvements in sensitivity of blood measures to assess trace mineral status await determination of the most critical metalloenzyme activity. At present, metalloenzyme activities are seldom more useful than concentrations of minerals in plasma and often are impractical because of loss of activity in shipment to the laboratory. Perhaps the ultimate assessment tool is the response of animals to supplementation.
Article
Full-text available
The rumen is the site of significant interactions between Cu, S and Mo. It also shows reactions between Cu, S and Fe. The interaction between Mo and S results in the formation of thiomolybdates, which in the absence of adequate quantities of rumen Cu are absorbed into the animal and bind to Cu in biological compounds. This is the cause of thiomolybdate toxicity, often misleadingly called Cu deficiency. The effects of thiomolybdates being absorbed into the animal are considered, especially how thiomolybdates bind to Cu-containing compounds such as enzymes, decreasing their activity without removing the active Cu component. The sources of Cu, Mo, Fe and S are examined, including the impacts of water and soil on the animal's intake. Within the present review we have been able to provide evidence that: all classes of thiomolybdates are formed in the rumen; in the absence of available Cu all thiomolybdates can be absorbed into the animal rapidly though the rumen wall or via the small intestine; thiomolybdates bind to Cu in biological compounds and are able to cause problems; effects of thiomolybdate are reversible in vivo and in vitro on cessation of thiomolybdate challenge; the tetra-thiomolybdate form is the most potent Cu binder with decreased potency with decreasing S in the compound. Fe will exacerbate a thiomolybdate problem but will not directly cause it.
Chapter
This 4th edition of the book entitled "Mineral Nutrition of Livestock" consists of 20 chapters dealing with the following topics: the requirement for mineral, natural sources of minerals, assessing and controlling mineral status in livestock, calcium, magnesium, phosphorus, potassium, sodium and chloride, sulfur, cobalt, copper, iodine, iron, manganese, selenium, zinc, occassionaly beneficial elements, potentially toxic elements, design of supplementation trials for assessing mineral deprivation, and minerals and humans. The book consists of 595 pages with a useful appendix and index.
Article
The inhibition of growth is a cardinal symptom of zinc deficiency. In animals fed a zinc-inadequate diet, both food intake and growth are reduced within 4–5 d. Despite the concomitant reduction in food intake and growth, reduced energy intake is not the limiting factor in growth, because force-feeding a zinc-inadequate diet to animals fails to maintain growth. Hence, food intake and growth appear to be regulated by zinc through independent, although well coordinated, mechanisms. Despite the long-term study of zinc metabolism, the first limiting role of zinc in cell proliferation remains undefined. Zinc participates in the regulation of cell proliferation in several ways; it is essential to enzyme systems that influence cell division and proliferation. Removing zinc from the extracellular milieu results in decreased activity of deoxythymidine kinase and reduced levels of adenosine(5′)tetraphosphate(5′)-adenosine. Hence, zinc may directly regulate DNA synthesis through these systems. Zinc also influences hormonal regulation of cell division. Specifically, the pituitary growth hormone (GH)–insulin-like growth factor-I (IGF-I) axis is responsive to zinc status. Both increased and decreased circulating concentrations of GH have been observed in zinc deficiency, although circulating IGF-I concentrations are consistently decreased. However, growth failure is not reversed by maintaining either GH or IGF-I levels through exogenous administration, which suggests the defect occurs in hormone signaling. Zinc appears to be essential for IGF-I induction of cell proliferation; the site of regulation is postreceptor binding. Overall, the evidence suggests that reduced zinc availability affects membrane signaling systems and intracellular second messengers that coordinate cell proliferation in response to IGF-I.
Article
The fourth edition of this important book covers the advances in livestock mineral nutrition, updated with more illustrations and additional material on the relationship between livestock and man. Recent developments are discussed, such as increasing the 'mineral value' of feeds by the use of additives and enhancing mineral availability through the use of organic sources of trace elements. The concept of the 'mineral footprint' of livestock production is introduced and methods of mineral feeding that lower environmental pollution are presented. Opportunities and problems in manipulating the mineral content of livestock to improve the mineral status of consumers are also addressed. The book is an essential resource for researchers and students in animal nutrition, agriculture and veterinary medicine, and a useful reference for those concerned with human nutrition and environmental protection.
Article
An automated soil test for sulphur and interpretative criteria for Saskatchewan conditions are presented. Sulphate‐sulphur (SO4‐S) in 0.01 M CaCl2 extracts of soil is determined by turbidimetric and nephelometric procedures operating simultaneously in an AutoAnalyzer system. A procedure for the segregation and elimination of samples containing excessive amounts of SO4‐S is described.The total amounts of soluble SO4‐S in the 0–60 cm soil layer is considered for interpretative purposes. Tentative critical values for two broad groups of field crops and a preliminary summary of the sulphur status of Saskatchewan soils based on analysis of farm samples are discussed.
Article
Although it has been known for a long time that hypocuprosis affects cattle in the province of Buenos Aires (Argentine), mostly during the spring, there has been no attempt to estimate the occurrence of this deficiency. A survey involving more than 200 farms was carried out in five districts of the Salado river basin, where hypocuprosis is endemic. A total of 4750 plasma samples were obtained during 1993 to 1996, and the Cu content was analyzed by atomic absorption spectrophotometry. Fifty-six per cent of cattle population in the five districts was hypocupremic (plasma Cu<60 μg dl−1), while 14% was severely hypocupremic (<29 μg dl−1). The highest incidence was during 1994, when 71% of cattle was hypocupremic and 26% was severely hypocupremic. During the spring a maximum of 37% of cattle was severely hypocupremic, which agrees with previous findings. The causes of hypocuprosis have been defined for the area, as low forage Cu with high S and Fe mainly during the autumn. A positive association between the occurrence of hypocuprosis and excesses of rainfall was found, that explain the seasonal pattern of hypocuprosis and the influence of floodings on hypocuprosis.
Article
A field survey involving 13 farms was carried out during the spring of 1994 in the Magdalena district to elucidate the causes of cattle hypocuprosis, that has been previously detected as a widespread condition in the area. From each farm, 30 blood samples and representative forage samples were obtained. Liver samples (N: 40) were obtained at a local slaughterhouse. Plasma and liver Cu, and forage Cu, Fe, Mo and S contents were analyzed. Plasma Cu concentrations were below 60 μg/dl in 78.6%, 37.6% and 61.9% of samples from calves, heifers and cows, respectively, in a total of 390 samples. Liver Cu concentration was below 75 ppm DM in 77.5% of the samples. The forage Cu concentration in 44 samples averaged 6.0 ppm DM, and was below the recommended level of 8 ppm in 84% of the samples. Molybdenum and S concentrations averaged 0.65 ppm DM and 0.28% DM, respectively. It is concluded that the widespread hypocuprosis develops as a simple deficiency, complicated by the high levels of S in the forages. This contrasts the Mo toxicity problems reported for other districts within the same basin.
Article
To characterize serum copper status of cows and heifers in beef cow-calf herds throughout the United States and to evaluate use of copper supplements in those herds. Cross-sectional survey. 2,007 cows and heifers from 256 herds in 18 states. Producers participating in a health and management survey conducted as part of the National Animal Health Monitoring System voluntarily allowed serum samples to be obtained from cows and heifers for determination of copper concentration. Results were categorized as deficient, marginally deficient, or adequate. The proportion of cattle and herds (on the basis of mean value of the tested cattle) in each category was determined. Copper concentrations were compared between herds that reportedly used copper supplements and those that did not. Overall, 34 of 2,007 (1.7%) cows and heifers were deficient in copper, and 781 (38.9%) were marginally deficient. In each region, at least a third of the cattle were deficient or marginally deficient. For herds, 92 of 256 (35.9%) were marginally deficient, and 22 (0.8%) were deficient. Approximately half of the producers reported use of copper supplements, but a sizeable proportion of those producers' cattle and herds were classified as marginally deficient or deficient. Copper deficiency is not restricted to a single geographic region of the United States. Copper deficiency can persist despite reported use of supplements by producers. Veterinarians dealing with beef cow-calf herds that have problems consistent with copper deficiency should not rule out copper deficiency solely on the basis of geographic region or reported use of copper supplements for the herd.