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

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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.
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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)
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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
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This study aims to reduce embryonic mortality, increase body weight, and improve immune system in chicken. A total of 240 eggs were assigned to three treatments (n = 60) and injected with cooper (Cu), zinc (Zn), and iron (Fe) loaded by montmorillonite (Mnt), and one untreated group (n = 60). Some hormones and enzymes related with growth were measured in terms of serum, and expression of some genes related to growth, immune, and programmed cell deaths that were determined in the liver and spleen of chicken by RT-qPCR. The embryonic death on the fifth and seventh days after injecting eggs with Fe-Mnt was less obvious than in other groups. The heaviest body weight was recorded for Fe-Mnt and Cu-Mnt treatment. Fe-Mnt treatment had higher serum GSH, SOD, GH, and Myostatin contents and lower MDA than those in the other treatments. Cu-Mnt treatment included the highest contents of CAT enzyme and IGF-1 hormone in serum. The highest expression of IGF-1, GH, BCL6, and SYK genes in liver tissue were recorded by Zn-Mnt, IGFBP2, FGF8, and IFNW1 genes by Cu-Mnt, and TC1RG1 and IFNW1 genes by Fe-Mnt in spleen tissue. In conclusion, Fe-Mnt was the best treatment for reducing embryonic mortality, and increasing body weight of chickens and expression of growth and immune genes, followed by Cu-Mnt treatment.
... Trace elements, especially copper (Cu) and zinc (Zn), have vital structural or catalytic roles in many metal-binding proteins and metalloenzymes, which are important for immune system function, nutrient metabolism [1][2][3], and health in rabbits; in addition, they had proven significant antimicrobial effect [4][5][6]. Pharmacological amounts of Zn and Cu, either incorporated into the diet or injected into the body, enhanced the growth performance of rabbits, particularly after/during the weaning stage [7]. The various sources of supplemental trace minerals and particularly Zn and Cu, such as oxide, nanoparticles, carrier loading, and ligand with complexes, showed different effects on animals' growth rate; some sources have a poor mineral bioavailability, leading to irritation in the intestinal mucosa or increase in the excretion of trace minerals to the intestinal environment [8]. ...
... The liver, in particular, is the major storage organ of Cu and Zn, and the stored Cu is largely bound to the metallothionein in most species, which have the capacity to bind both physiological (such as zinc, copper, and selenium) and xenobiotic (such as cadmium, mercury, silver, and arsenic) heavy metals through the thiol group of its cysteine residues [44,45]. The blood biochemical variables of calves were improved under Zn and Cu administration [6,46]. ...
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Simple Summary: Zinc and Copper are two important trace minerals, which are involved in numerous vital biological activities in animal's body, such as enzyme activation and enhancement of immune function, growth, reproduction, DNA synthesis, cell division, and neurotransmitters production. Recently, the application of trace mineral organic complexes in animal feed received much more attention than the inorganic sources. The organic sources can contribute significantly to improving animals' health and reproduction, as the minerals are more biologically available and absorbable than they are when coming from the inorganic sources. In this study, three injectable varieties of Zn and Cu supplementation, sulfate, inorganic (loaded with montmorillonite), and novel organic (ligand with triazine hydrazone), were tested with weaned rabbits. The results revealed that these three mineral types vary at the most biological responses, and only one category of our novel organic complexes provided consistent animal performance improvement, including weight gain, serum antioxidant, meat quality, intestine morphometry, and the expression of peptide growth factors and cytokine genes. To our knowledge, this is the first work on the Zn and Cu with triazine hydrazone ligand as two organic complexes in rabbits. Abstract: Two novel transitional organic Zn/Cu complexes based on a new biocompatible bidentate triazine-hydrazone ligand (Thz) was designed, synthesized, and evaluated in this study. This study evaluated the effects of injecting 60 mg of Zn and 40 mg of Cu in three different forms, twice per week, for eight weeks on growth performance, expression of growth factors and cytokine genes, carcass yield, blood biochemicals, and intestinal morphology in weaned rabbits. The tested complexes were sulfate (Cu/ZnSO4), montmorillonite (Cu/Zn-Mnt), and triazine hydrazone (Cu/Zn-Animals 2019, 9, 1134 2 of 17 Thz). A total of 60 V-line weaned rabbits at four weeks of age were assigned to four treatments (n = 15), which were intramuscularly injected with 0.5 mL of either (1) saline (control) or saline containing (2) Cu/ZnSO4, (3) Cu/Zn-Mnt, or (4) Cu/Zn-Thz. Compared to the controls, the rabbits injected with Cu/Zn-Thz showed a higher (p < 0.01) growth rate, carcass yield (p < 0.05), and liver expression of insulin like growth factor-1 (IGF-1), growth hormone receptor (GHR), fibroblast growth factor-1 (FGF1), and transforming growth factor beta-1 (TGFB1) (p < 0.05), as well as better jejunum morphometric variables (p < 0.05). On the other hand, mRNA of FGF1, TGF1, TCIRG1, and adenosine deaminase (ADA) were higher expressed (p < 0.05) in the spleen tissues of Cu/Zn-Mnt group. Collectively, the results indicated that our novel synthesized organic complexes of Zn/Cu-Thz proved to be a suitable feed supplement, as it increased rabbit productive performance through enhancing expression of peptide growth factors and cytokine genes.
... Trace elements, especially copper (Cu) and zinc (Zn), have vital structural or catalytic roles in many metal-binding proteins and metalloenzymes, which are important for immune system function, nutrient metabolism [1][2][3], and health in rabbits; in addition, they had proven significant antimicrobial effect [4][5][6]. Pharmacological amounts of Zn and Cu, either incorporated into the diet or injected into the body, enhanced the growth performance of rabbits, particularly after/during the weaning stage [7]. The various sources of supplemental trace minerals and particularly Zn and Cu, such as oxide, nanoparticles, carrier loading, and ligand with complexes, showed different effects on animals' growth rate; some sources have a poor mineral bioavailability, leading to irritation in the intestinal mucosa or increase in the excretion of trace minerals to the intestinal environment [8]. ...
... The liver, in particular, is the major storage organ of Cu and Zn, and the stored Cu is largely bound to the metallothionein in most species, which have the capacity to bind both physiological (such as zinc, copper, and selenium) and xenobiotic (such as cadmium, mercury, silver, and arsenic) heavy metals through the thiol group of its cysteine residues [44,45]. The blood biochemical variables of calves were improved under Zn and Cu administration [6,46]. ...
Article
Full-text available
Two novel transitional organic Zn/Cu complexes based on a new biocompatible bidentate triazine-hydrazone ligand (Thz) was designed, synthesized, and evaluated in this study. This study evaluated the effects of injecting 60 mg of Zn and 40 mg of Cu in three different forms, twice per week, for eight weeks on growth performance, expression of growth factors and cytokine genes, carcass yield, blood biochemicals, and intestinal morphology in weaned rabbits. The tested complexes were sulfate (Cu/ZnSO4), montmorillonite (Cu/Zn-Mnt), and triazine hydrazone (Cu/Zn-Thz). A total of 60 V-line weaned rabbits at four weeks of age were assigned to four treatments (n = 15), which were intramuscularly injected with 0.5 mL of either (1) saline (control) or saline containing (2) Cu/ZnSO4, (3) Cu/Zn-Mnt, or (4) Cu/Zn-Thz. Compared to the controls, the rabbits injected with Cu/Zn-Thz showed a higher (p < 0.01) growth rate, carcass yield (p < 0.05), and liver expression of insulin like growth factor-1 (IGF-1), growth hormone receptor (GHR), fibroblast growth factor-1 (FGF1), and transforming growth factor beta-1 (TGFB1) (p < 0.05), as well as better jejunum morphometric variables (p < 0.05). On the other hand, mRNA of FGF1, TGF1, TCIRG1, and adenosine deaminase (ADA) were higher expressed (p < 0.05) in the spleen tissues of Cu/Zn-Mnt group. Collectively, the results indicated that our novel synthesized organic complexes of Zn/Cu-Thz proved to be a suitable feed supplement, as it increased rabbit productive performance through enhancing expression of peptide growth factors and cytokine genes.
... 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). ...
... 8 In addition to its biological functions, Cu was already investigated as a growth enhancer for several animal species. [11][12][13] Indeed, earlier studies have reported that Cu supplementation with a 30-300 mg/ kg diet increases growth indicators and feed efficiency in rabbits. [14][15][16] Different sources of Cu such as sulfate, chloride, oxide, methionine, proteinate, acetate and loaded with montmorillonite or triazine hydrazone have been applied in rabbit diets. ...
Article
The various sources of supplemented copper had a different impact on the animal performance based on their bioavailability. The current study compared the effects of supplementary copper oxide (CuO), copper acetate (Cu-acetate) and copper nanoparticles (Cu-NP) on performance, immune function, nutrients digestibility and architecture of the liver and kidney of growing rabbits for eight weeks. Sixty rabbits (581 ± 6.56 g) were randomly allocated to four treatments as follows: basal diet, 100 mg copper/kg diet as CuO, 100 mg copper/kg diet as Cu-acetate and 50 mg copper/kg diet as Cu-NP. Cu-acetate and Cu-NP improved specific growth rate, final weight and daily weight gain. Cu-NP supplementation had higher feed intake, feed conversion, protein efficiency, hematocrit and hemoglobin values compared with other copper forms. All copper sources showed higher levels of serum complement component 3, Immunoglobulin M, lysozyme activity and the digestibility of nitrogen-free extract, dry matter and organic matter. As a result, increased nutritive values were detected when the rabbits were fed copper-supplemented diets. No liver and kidney architecture alterations were identified between the experimental groups. In conclusion, both dietary Cu-NP and Cu-acetate were more efficient than CuO in enhancing growth and seem promising in fattening rabbit nutrition.
... 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. [31] and Mattiolli et al. [32], in which steers that received copper supplementation up to the maximum recommended level showed no change in body mass gain. ...
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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. 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, 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.
... Also, changes in the concentrations of acute phase response and inflammatory mediators make the transition cows susceptible to a variety of diseases including mastitis [7]. Moreover, several host immune functions such as phagocytosis, antibody development, and cytokine production get suppressed during the transition period [8].The health status of newborn calves is one of the most important animal health issues facing the livestock industry and safeguarding its well-being is equally essential as today's calves are known to be tomorrow's herd [9,10]. The newborn calves cannot fight the infectious agents as their immune system is underdeveloped at birth. ...
Article
Background Periparturient period induces stress in cows which fluctuates hormonal and metabolic function and causes immune suppression. Apart from impairing the health, production, and reproduction of cows, it also influences the well-being of newborn calves by decreasing the colostrum quality. Micronutrients are known for optimal health and production and their effects on parturition stress, immune response in both cow and its calf need to be explored. Aim The aim of this study was to see the effect of oral supplementation of micronutrients during the prepartum period on the health status of crossbred dairy cows and subsequently on their newborn calves. Methods A total of 42 healthy multiparous cows were selected and randomly divided into five groups with seven cows in each group, i.e. control (Basal Diet, BD), VA group (BD + vitamin A, 10⁵ IU), Zn group (BD + zinc sulphate, 60 ppm), VE group (BD + vitamin E, 2500 IU), and combined supplementation (CS) group (BD + combination of VA, Zn, and VE). The supplements were offered in compounded concentrate DM (100 g) to individual cows once daily before the morning feeding and the remaining portion was incorporated in the TMR. Feeding was started one month before the expected days of calving till calving. Blood samples were collected from cows at days -15, -7, -3, 0, +3, +7, and +15 relative to the day of calving. Blood samples from newborn calves and milk samples of cows were collected at days 0, +3, +7, and +15. Milk somatic cell counts (SCC) were estimated using a cell counter. Cortisol was estimated by ELISA kit in blood and milk plasma of cows and in the blood plasma of their calves. Total immunoglobulins (Ig) were estimated in milk of cows and serum of calves using zinc sulphate turbidity method. Blood neutrophils from cows and calves were studied for phagocytic activity (PA) using nitro blue tetrazolium (NBT) assay.Data were analysed by repeated-measures two-way ANOVA using the mixed procedure of SAS, and the pairwise comparison was performed using a multiple comparison test (Tukey). Results Combined supplementation of micronutrients decreased (P < 0.05) maternal blood plasma (control vs. CS group, 5.98 ± 0.20 vs. 3.86 ± 0.23 ng/ml) and milk plasma (3.96 ± 0.13 vs. 2.71 ± 0.10 ng/ml) cortisol, milk SCC (3.05 ± 0.11 vs. 2.12 ± 0.10 × 10⁵ cells/ml) and increased (P < 0.05) total milk Ig concentration (18.80 ± 0.11 vs. 23.04 ± 0.57 mg/ml) and the PA of blood neutrophils (0.84 ± 0.03 vs. 1.07 ± 0.03). Similarly, lower blood cortisol concentration (9.69 ± 0.35 vs. 6.02 ± 0.18 ng/ml) and higher (P < 0.05) total Ig (23.26 ± 0.11 vs. 30.34 ± 0.70 mg/ml) and PA of blood neutrophils (0.37 ± 0.02 vs. 0.52 ± 0.02) were observed in the calves born to CS group of cows as compared to the control. Highest (P < 0.05) positive effects (lower stress levels and higher immune response) of treatment were noticed in CS group followed by VE group and then Zn group. However, VA group didn’t differ from the control group. Conclusion Our results indicate that micronutrient interventions during the prepartum period can improve the health status of dairy calves and subsequently the well-being of their calves.
Article
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.
Article
Neonatal diarrhea in dairy calves causes huge economic and productivity losses in the dairy industry. Zinc is an effective anti-diarrheal agent, but high doses may pose a threat to the environment. Therefore, we aimed to evaluate the effects of low-dose zinc supplementation on the growth, incidence of diarrhea, immune function, and rectal microbiota of newborn Holstein dairy calves. Thirty newborn calves were allocated to either a control group (without extra zinc supplementation), or groups supplemented with either 104 mg of zinc oxide (ZnO, equivalent to 80 mg of zinc/d) or 457 mg of zinc methionine (Zn-Met, equivalent to 80 mg of zinc/d) and studied them for 14 d. The rectal contents were sampled on d 1, 3, 7, and 14, and blood samples were collected at the end of the study. Supplementation with ZnO reduced the incidence of diarrhea during the first 3 d of life, and increased serum IgG and IgM concentrations. The Zn-Met supplementation increased growth performance and reduced the incidence of diarrhea during the first 14 d after birth. The results of fecal microbiota analysis showed that Firmicutes and Proteobacteria were the predominant phyla, and Escherichia and Bacteroides were the dominant genera in the recta of the calves. As the calves grew older, rectal microbial diversity and composition significantly evolved. In addition, dietary supplementation with ZnO reduced the relative abundance of Proteobacteria in 1-d-old calves, and increased that of Bacteroidetes, Lactobacillus, and Fecalibacterium in 7-d-old calves, compared with the control group. Supplementation with Zn-Met increased the relative abundance of the phylum Actinobacteria and the genera Fecalibacterium and Collinsella on d 7, and that of the genus Ruminococcus after 2 wk, compared with the control group. Thus, the rectal microbial composition was not affected by zinc supplementation but significantly evolved during the calves' early life. Zinc supplementation reduced the incidence of diarrhea in young calves. In view of their differing effects, we recommend ZnO supplementation for dairy calves during their first 3 d of life and Zn-Met supplementation for the subsequent period. These findings suggest that zinc supplementation may be an alternative to antibacterial agents for the treatment of newborn calf diarrhea.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
Article
Zinc (Zn) administration at non-toxic doses protects against the hepatotoxicity produced by many agents, but the underlying mechanisms remain elusive. Aim: To examine the basis of Zn-induced generalised hepatoprotective effects. Rats and mice were given Zn at known hepatoprotective levels (100 mumol ZnCl2/kg/day, s.c., for 4 days) and molecular responses were assessed. Zn treatment produced changes in 5% of the genes on custom-designed mouse liver array and Rat Toxicology-II array. Changes in gene expression were further confirmed and extended by real-time reverse transcriptase-polymerase chain reaction. Zn treatment dramatically increased the expression of the metallothionein (Mt), and modestly increased the expression of acute-phase protein genes (ceruloplasmin, Stat3, egr1, Cxc chemokines and heat-shock proteins). For genes encoding for antioxidant enzymes, some were increased (Nrf2 and Nqo1), while others remained unaltered (Cu, Zn SOD and glutathione S-transferases). Expressions of cytokine and pro-inflammatory genes were not affected, while genes related to cell proliferation (cyclin D1) were modestly upregulated. Some metabolic enzyme genes, including cytochrome P450s and UDP-glucuronosyltransferase, were modestly suppressed, perhaps to switch cellular metabolic energy to acute-phase responses. Liver Zn content was increased between 1.6- and 2.1-fold, while hepatic MT protein was increased between 50 and 200-fold. Mice typically showed greater responses than rats. Such gene expression changes, particularly the dramatic induction of MT and Nrf2 antioxidant pathway, occur in the absence of overt liver injury, and are probably important in the hepatoprotective effects of Zn against toxic insults.